ICS 07.080  
                                                                                                                    AFDC 11 (12 90) DTZS  
      
                               DRAFT TANZANIA STANDARD                                                                          
 
Biotechnology - Biobanking - Process and quality requirements for 
establishment, maintenance and characterization of mammalian cell lines  
 
 
 
 
 
 
 
 
 
 
 
 
 
                            TANZANIA BUREAU OF STANDARDS   
  
 0 foreword  
Scientific research using cell lines has contributed greatly to the understanding of human health. Cell 
cultures are increasingly used to complement studies using animal models . Although cell lines are 
important research tools, potential problems have recently been identified.  
Cell lines have unique characteristics and behavior  that can change as they continue to be passaged. 
The original phenotype (e.g. expression of specific b iomarkers) can be lost or new characteristics or 
behavior  (e.g. development of tumorigenicity) may develop. It is important to minimize passaging to retain 
the original characteristics that were present when the cell line was first established.  
Other probl ems such as contamination, either with microorganisms or another cell line, and 
misidentification can also arise. Cultures can become contaminated during cell line establishment or later 
when cultures are passaged. These problems are often not visible by e ye and require specific testing to 
be detected.  
This Tanzania standar d was developed in order to  address the above mentioned issues and provides  
technical specifications for biobanks that handle mammalian cell lines.  
 
During the development of this Tanzan ia standard assistance was drawn from the following publication;  
 ISO 21709:2020  Biotechnology - Biobanking - Process and quality requirements for 
establishment, maintenance and characterization of mammalian cell lines  
 ISO 21709:2020/Amd.1:2021  Biotechnolog y - Biobanking - Process and quality requirements for 
establishment, maintenance and characterization of mammalian cell lines AMENDMENT 1 , 
Published by International Organization for Standardization (ISO).  
In reporting the results of a test or analysis mad e in accordance with this draft Tanzania Standard, if the 
final value observed or calculated is to be rounded off, it shall be done in accordance with TZS 4 (see 
clause 2).  
1 Scope  
This Tanzania standard specifies process and quality requirements for the b iobanking of mammalian 
(including human) cell lines. It describes requirements for the fundamental procedures of the biobank 
handling cell lines, such as establishment, reception, identification, propagation, preservation, storage, 
quality control, and dis tribution of cell lines.  
This Tanzania standard  can be used by organizations performing biobanking activities with mammalian 
cell lines used for research and development, biobank users, organizations and schemes using peer -
assessment and accreditation bodi es. 
This Tanzania standard  does not apply to biological material intended for therapeutic use.  
NOTE : National regulations or requirements can also apply to specific topics covered in this Tanzania 
standards . 
2 Normative references  
The following referenced documents are indispensable for the application of this standard. For dated 
references, only the edition cited applies. For undated references, the latest edition of the referenced 
document (including any amendments) applies;  
TZS 4 Rounding off numerical v alues  
TZS 2973/ ISO 20395  Biotechnology - Requirements for evaluating the performance of quantification 
methods for nucleic acid target sequences - qPCR and dPCR .  
 TZS 2974 -1/ISO 20391 -1 Biotechnology - Cell counting - Part 1: General guidance on cell counti ng 
methods.  
TZS 2974 -2/ISO 20391 -2 Biotechnology - Cell counting - Part 2: Experimental design and statistical 
analysis to quantify counting method performance  
TZS 2975 /ISO 20387  Biotechnology - Biobanking - General requirements for biobanking .  
3 Terms a nd definitions  
For the purposes of this draft Tanzania standard , the following terms and definitions apply.  
3.1 cell culture  
growth of cells dissociated from the parent tissue by spontaneous migration or mechanical or enzymatic 
dispersal for propagation an d consecutive passages in vitro  
3.2 Cell line  
Progeny  of a primary culture (3.16) after it has been passaged beyond crises or beyond senescence 
either spontaneously or after introduction of immortalizing factors  
Note: A cell line is continuous for prolifer ation. 
3.3 Cell morphology  
Form  and structure of the cell . 
Note:  Morphology can be represented by a single parameter or a combination of two or more parameters.  
3.4 Cell strain  
Progeny  of a primary culture (3.16) before it has been passaged beyond crises o r beyond senescence or 
immortalized by introduction of immortalizing factors  
Note:  Not all the cell strains will continue proliferation to reach the cell lines . 
3.5 Cell type  
Classification  used to distinguish among distinct cell forms  
3.6 Cryopreservation  
Process  by which cells are maintained in in an ultra -low temperature (e.g. − 40 °C to − 70 °C) at an 
inactive state so they can be revived later  
3.7 Derivative material  
Biological  material that was derived from the original donation  
EXAMPLE : Derivative mate rials can be cell lines.  
3.8 Doubling time  
Time  taken for cultured cell count to double  
3.9 Establishment of cell line  
Process  of producing a cell line capable of indefinite proliferation  
3.10 Identity verification   
 Part of the process of verifying authenti city of a cell line in which cell origin is genetically confirmed  
3.11 Informed consent  
Process by which an individual or its designated legal representative or nominated representative 
voluntarily confirms willingness to donate biological material for res earch purposes, after having been 
informed of all aspects of the potential research that are relevant for the decision to donate  
3.12 M icrobe/ microorganism  
Unicellular  living cells which cannot be observed with naked eye but only under a microscope  
3.13 Passage number  
Number  of sub culturing  that occurred  
3.14 Population doubling level ( PDL) 
Total  number of population doublings of a cell line (3.2) or cell strain (3.4) since its initiation in vitro  
3.15 Primary cells  
Cells  isolated directly from tissue or o rgans taken directly from an organism, using enzymatic or 
mechanical methods  
3.16 Primary culture  
Culture  started from cells, tissues, or organs taken directly from an organism, and before the first 
subculture, propagation and consecutive passages in vitro  
3.17 Viability  
Attribute  of being alive (e.g. metabolically active, capable of reproducing, have intact cell membrane, or 
have the capacity to resume these functions) as defined based on the intended use  
4 Requirements  
4.1 General  
4.1.1  During the whole c ell culture workflow, precautions shall be taken to avoid cross contamination 
between different samples/specimens, e.g. by using disposable material whenever feasible or appropriate 
cleaning procedures between processing of different specimens/samples.  TZS 2975 , Clause 4, shall be 
followed.  
4.1.2  The biobank shall establish, implement, and maintain a quality management system as described in 
TZS 2975 , Clause 8.  
4.2 Legal and ethical requirements  
4.2.1  TZS 2975 , 4.3 shall be followed.  
4.2.2  The biobank shall  consult a competent institutional/independent ethics review committee, which is 
responsible for the investigation and evaluation of any related ethical principles/requirements.  
4.2.3  The biobank shall comply with TZS 2975 , 4.1.6 and 7.2.3.4. The biologica l material, which is the 
source of the cell line, shall be collected, transported and handled according to internationally accepted 
procedures.   
 4.2.4  The biobank shall be aware of and able to demonstrate compliance with relevant national, regional 
and inte rnational approved ethics, laws and regulations relating to the biological material held in the 
biobank.  
4.2.5  Further ethical requirements related to the informed consent are included in 4.6.  
4.2.6  The biobank shall meet the organizational structure, reso urce, process, and quality requirements 
described in TZS 2975 , Clauses 5, 6 and 7, in addition to those in this document.  
4.3 Facilities  
4.3.1 General  
4.3.1.1  TZS 2975 , 6.3 shall be followed . 
4.3.1.2  The processing facility shall be constructed and operate d to minimize the introduction, generation, 
and retention of particles and microorganisms.  
4.3.1.3  The biobank shall define a protocol to test the quality of the biobanking environment(s).  
4.3.2 Cell culture facility  
4.3.2.1  The biobank culturing and prese rving cell lines shall establish a cell culture facility.  
4.3.2.2  The biobank shall assess the biological safety level of the cell lines that will be handled. The 
biobank shall assess the risk of asphyxiation based on the number and condition of liquid nit rogen tanks 
and storage methods and install an oxygen monitor accordingly.  
4.3.2.3  The biobank shall meet biological safety and security requirements for its facilities to minimize the 
risk to personnel and surroundings from handling the cell lines.  
4.3.2. 4 The biobank shall take necessary measures to control airflow.  
4.3.2.5  The biobank should make efforts to secure a separate space for storage of duplicated cell lines in 
order to prevent the loss of collections in emergency situations. If the biobank cann ot secure a separate 
alternate space for storage, the biobank should duplicate cell lines and store them in separate liquid 
nitrogen tanks. If the biobank cannot duplicate all stored cell lines, the biobank should make duplicates of 
the relevant cell lines  or those that can be required by contractual agreements.  
4.3.2.6  The biobank shall designate a separate area for handling deposited cell lines and/or tissues until 
they are determined to be free of microbial contamination.  
4.4 Equipment  
4.4.1 General  
4.4.1.1 TZS 2975 , 6.5 shall be followed.  
4.4.1.2  Equipment used in biobanking should be selected under quality specific criteria defined by the 
biobank, and for managing risks that can affect or impact performed assays, tests or their validity.  
4.4.1.3  All inc ubators used, for working in accordance with this document, shall be monitored.  
4.4.1.4  The biobank shall be furnished with or have controlled access to the following equipment as 
minimum requirements for cell culturing and ensure quality and safety requir ements regarding the 
equipment.  
a) Biological safety cabinet (Class II, Class III): physical structure protecting cell cultures from 
contaminants and personnel from potentially harmful biological substances. The cabinet can require  
 appropriate biosafety me asures, depending upon the biosafety level of the biological materials being 
handled.  
b) Cell counting equipment: an automated cell counting feature is recommended, manual cell counting 
slide (e.g. hemocytometer slide) may also be used.  
c) CO2 incubator: a  self-cleaning feature is recommended;  
d) Freezer: data logger, alarming system and backup system are recommended.  
e) Refrigerator: data logger, alarming system and backup system are recommended.  
f) Liquid nitrogen tank for vapor/liquid phase storage of ce ll lines with temperature monitor and alarming 
system are recommended.  
g) microscope: a camera for documentation is recommended;  
h) Other general laboratory equipment (e.g. autoclave, deep freezer, refrigerator, centrifuge).  
i) Inverted microscope (equippe d with a camera).  
4.4.1.5  The biobank should be furnished with or have controlled access to the equipment used for 
controlled rate freezing of cell lines.  
4.4.1. 6 The biobank shall equip its facility with appropriate biosafety equipment based on the assess ed 
biological safety level (documented risk analysis); see 4.3.2. TZS 2975 , 6.3.5, shall be followed.  
4.4.1. 7 The biobank shall have personal protective equipment available in accordance with the biosafety 
level of the facility and the nature of the cell l ine. Personnel within the facility shall wear appropriate 
personal protective equipment during cell culturing to prevent contact with culture media or aerosol that 
can be produced during the process.  
4.4.1.8  Personnel handling frozen materials or liquid ni trogen shall use cryogenic personal protective 
equipment.  
4.4.1.9  Storage equipment for cell lines should have a monitoring system and an alarming device or 
remote control system for immediate response to equipment malfunction or breakdown. The biobank sha ll 
keep records of equipment malfunction or breakdown and appropriate follow -up measures. The biobank 
should have an emergency generator or other equipment necessary to protect cell lines when power is 
lost. 
4.4.1.10  The biobank should have the necessary b ackup equipment (e.g. vacant gas CO2 incubators, 
freezers, and liquid nitrogen tanks) available in the case of equipment malfunction or failure.  
4.4.2 Equipment inspection  
4.4.2.1  The biobank shall inspect at planned intervals the following pieces of equip ment, and their 
characteristics, used for cell lines, when applicable:  
a) autoclave: according to the safety level, pressure, temperature, safety valve, steam outlet, cleaning 
and microbiological and chemical control of the process;  
b) biological safety ca binet: airflow, HEPA filter condition, sterility and cleaning;  
c) CO2 incubator: temperature, humidity, CO2 system, sterility and cleaning;  
d) liquid nitrogen tank: liquid nitrogen level, temperature and inspection of alarming system;  
e) microscope: compon ents and cleaning;   
 f) refrigerator: temperature and stability of electric supply;  
g) ultra -low temperature freezer: status of compressor, temperature system, and stability of electric 
supply;  
h) ventilation system: inspection;  
i) oxygen monitor devices: in spection;  
j) backup equipment: status and stability of electrical supply and/or supply of nitrogen or CO2 at 
emergency of electricity failure.  
4.4.2.2  Inspection records shall be retained. Details of any deviations shall be documented.  
4.5 Reagents  
4.5.1 Reagents in biobanking should be selected under quality specific criteria defined by the biobank, 
and for managing risks that can affect or impact performed assays, tests or their validity.  
4.5.2 The biobank should establish, document and implement a proced ure for evaluating the reagent 
stability; this procedure should define criteria for internal or external quality control (i.e. antibodies, 
diluents, calibrators).  
4.6 Informed consent  
4.6.1 General  
4.6.1.1  A procedure for obtaining the informed consent sha ll be established, documented and 
implemented. This informed consent procedure applies to human donors/patients and the responsible 
party for donor animals. In case of donations from animals, the donor's legally designated or nominated 
representative can b e understood as the animal’s responsible party.  
4.6.1. 2 The informed consent procedure shall comply with the requirements set by relevant ethics review 
committees. Appropriate approval of the procedure shall be obtained from the relevant ethical committee.  
Evidence of ethical approval shall be available.  
4.6.1. 3 The informed consent procedure shall take into account language barriers, condition and the 
educational level of the donor through the use of native non -technical language that is understandable to 
the donor/patient or the donor's/patient’s legally designated or nominated representative.  
NOTE In case the donor cannot understand the explanation, the responsible proxy can act on behalf.  
4.6.1 .4 During the informed consent procedure, it shall be ensured  that the donor/patient or the 
donor's/patient’s legally designated or nominated representative is informed about the aspects of his/her 
voluntary donation, and that these are understood.  
4.6.1. 5 Informed consent shall be obtained in writing from the donor /patient or the donor's/patient’s legally 
designated or nominated representative with a personally dated signature, and the informed consent 
process shall be finalized and documented before a donation of biological material is obtained.  
NOTE Dated signatur es can be written on paper, electronical or digital.  
4.6.1. 6 The informed consent form consists of an information form (see 4.6.4) and an informed consent 
signature form (see 4.6.5). These two forms can either be combined in one document or separated into 
two documents.  
4.6.2 Procedure of obtaining informed consent  
The biobank shall comply with the following elements while obtaining informed consent:   
 a) all aspects of the potential use are relevant to the donor’s/patient’s or the donor's/patient’s legally 
designated or nominated representative’s decision for the voluntary donation;  
b) the agreement on the informed consent shall not be affected by coercion, inducements, improper 
influences and/or misconduct;  
c) do not waive or appear to waive the donor’s/pati ent’s legal rights;  
d) use native non -technical language that is understandable to the donor/patient or the donor's/patient’s 
legally designated or nominated representative;  
e) provide ample time for the donor/patient or the donor's/patient’s legally desig nated or nominated 
representative to read and understand the informed consent form and to consider the voluntary donation;  
f) ensure personally dated signatures of the donor/patient or the donor's/patient’s legally designated or 
nominated representative an d the responsible physician or an authorized designee responsible for 
conducting the informed consent process;  
g) provide the donor/patient or the donor's/patient’s legally designated or nominated representative with a 
copy (in paper or digital) of the sig ned and dated informed consent form, which was approved by the 
biobank, and any other written information;  
h) ensure documentation of the process in the donor’s/patient’s source documents and maintaining the 
biobank’s signed informed consents with the esse ntial documents;  
i) show how informed consent will be obtained and recorded in special circumstances (4.6.3) where the 
donor/patient is unable to provide it him - or herself.  
4.6.3 Special circumstances for informed consent  
4.6.3.1 Donor/patient needing leg ally designated or nominated representatives  
Informed consent may be given by legally designated or nominated representatives only if a donor/patient 
is unable to make the decision to donate (e.g. infant, child, juvenile, seriously ill patient, donor/patie nt with 
a mental, intellectual disability, animal). In such cases, except for donor animals, the donor/patient shall 
also be informed about the potential research within his/her ability to understand.  
4.6.3. 2 Human donor/patient unable to read or write  
Informed consent shall be obtained through a supervised oral process, if a donor/patient or legally 
designated or nominated representative is unable to read or write. An independent and impartial witness 
shall be present throughout the process. The written in formed consent form and any other information 
shall be read aloud and explained to the donor/patient or his/her legally designated or nominated 
representative. Wherever possible, either the donor/patient or his/her legally designated or nominated 
represent ative shall sign and personally date the informed consent form. The witness shall sign and 
personally date the informed consent form attesting that the information was accurately explained, and 
that informed consent was freely given.  
4.6.4 Information prov ided to the donor/patient or the donor's/patient’s legally designated or 
nominated representative  
4.6.4.1  All information pertinent to potential use, including at least the following topics, shall be provided 
in writing and in native non -technical language  that is understandable to the donor/patient or the 
donor's/patient’s legally designated or nominated representative:  
a) potential research under appropriate conditions;  
b) potential use in many laboratories in many countries;   
 c) research applications;  
d) unexpected feedback (for example, the cells’ future use);  
e) potential commercial exploitation;  
f) other issues applicable to the research;  
g) statements regarding manipulation, retention and personal data protection.  
4.6.4.2  The informed consent form shal l make the donor/patient or the donor's/patient’s legally designated 
or nominated representative aware of and give the ability to accept or reject the donation based on the 
information of:  
a) the donation being a set of biological material and its associat ed data;  
b) that these cells will be stored, and may be sequenced, used in animal studies, or sent to other 
laboratories for analyses;  
c) the potential extent of testing to be performed on the tissue and/or the cells derived;  
d) that donations will be deid entified, thus either anonymized or pseudonymized/codified to maintain the 
donor's/patient’s confidentiality according to TZS 2975 , 4.3.  
4.6.4.3  The informed consent form and discussion should cover information that addresses:  
— key aspects of human cell r esearch; including but not limited to the fact that an immortal cell line can 
be established, which is a partial or full genetic match to the biomaterials donor;  
— that the cell line can be shared with other researchers outside the institution for other re search 
purposes that are not fully anticipated at this time.  
The informed consent form shall include a statement of possible consequences of the consent 
withdrawal. This shall include that withdrawal of consent means that derivative materials cannot be 
withdrawn.  
4.6.5 Informed consent signature  
The informed consent signature form shall contain the following:  
a) the agreement on the voluntary donation and to follow the responsible physician’s instructions;  
b) a statement declaring that refusal of donation i ncurs no penalty for the donor/patient;  
c) a statement declaring that the withdrawal of consent and thereby revoking the informed consent at any 
time incurs no penalty for the donor/patient;  
d) a statement with regard to the possible consequences of withdr awal;  
e) an acknowledgement of the information provided and confirmation that all the questions of the 
donor/patient or those of his/her legally designated or nominated representative were answered, that the 
donor/patient or his/her legally designated or n ominated representative acknowledges the information 
provided during the informed consent procedure and that (s)he had ample time to consider donating.  
f) a statement confirming that the donor/patient or his/her legally designated or nominated representati ve 
agrees to the use of the donor’s/patient’s relevant personal data for the purpose of research.  
4.7 Personnel  
4.7.1 General   
 4.7.1.1  The requirements of TZS 2975 , 6.2.1 apply.  
4.7.1.2  Vaccination of personnel is compulsory according to risk assessment.  
4.7.2 Personnel competence  
4.7.2.2  The requirements of TZS 2975 , 6.2.2 apply.  
4.7.2. 3 The biobank shall ensure that all personnel are knowledgeable about the health and safety risks 
of handling cell lines. The biobank shall provide safety training with regul ar updates based on the 
biosafety level of the facilities.  
4.7.2. 4 The biobank shall ensure that personnel involved in cell culturing are competent to perform the 
relevant tasks and knowledgeable in detection and prevention of microbial and cross contamina tion and 
misidentification.  
4.7.3 Personnel training  
4.7.3.1  The requirements of TZS 2975 , 6.2.3 apply.  
4.7.3. 2 The biobank shall provide biobank personnel (internal and/or external) with appropriate and 
relevant training with regular updates. The training  subjects include, but are not limited to the following:  
a) cell culture techniques, including aseptic laboratory skills;  
b) characterization and authentication of cell lines;  
c) establishment of cell lines;  
d) ethics and data confidentiality and security;  
e) health and safety;  
f) preservation and storage methods;  
g) environment control of cell culture areas;  
h) quality control of cell lines, including detection and prevention of microbial and cross contamination and 
misidentification;  
i) transport of cell lines, including e.g. relevant regulations in cell line shipment.  
4.7.3. 3 The biobank should periodically verify the knowledge of employees who manually perform the 
technological process to make sure that the process is carried out in accordance with inter nal procedure.  
4.7.4 Biorisk and biosafety of personnel  
The biobank shall assess biorisks of cell lines at the facilities and implement appropriate biosafety 
measures for the protection of personnel.  
5 Process requirements  
5.1 Establishment of cell lines w ithin the biobank  
5.1.1 General  
5.1.1.1  The requirements of TZS 2975 , 7.1 apply.  
5.1.1.2  The biobank can produce cell lines from cells or tissues derived from mammals, including 
humans.   
 5.1.1.3  The biological material which is the source of the cell line s hall be collected, transported, and 
handled according to existing Tanzania Standards  or internationally accepted procedures.  
5.1.1.4  The biobank shall obtain clinical and biological characteristics regarding the source of the cell line, 
including donor spe cies, origin, ethnic origin, sex, age, collected part (e.g. tissue, blood, etc.), and 
disease state, as feasible and appropriate. The biobank shall store all the information of to the 
established cell lines and record their preservation and storage informa tion. 
5.1.1.5  The biobank shall ensure that materials are collected and handled in accordance with relevant 
ethical requirements. The biobank shall handle the clinical information and other data regarding the 
donor/sources of the cells in such a way as to maintain security and confidentiality of the donor identity.  
5.1.1.6  The biobank shall provide a unique identifier to the cell line in an internationally recognized and 
distinct naming format in accordance with TZS 2975 , 7.5 a). Traceability shall be ensur ed according to 
TZS 2975 , 7.5.  
NOTE The biobank can access the International Cell Line Authentication Committee website  
(ICLAC: https:// iclac .org) or Cellosaurus (https:// web .expasy .org/ cellosaurus/ ) as references . 
5.1.1.7  The biobank shall establis h and implement measures to prevent cross contamination.  
5.1.1.8  The biobank shall implement correct aseptic technique while handling cell lines. The biobank 
should refrain from using antibiotics or antimycotic. If the biobank uses antibiotics or antimycot ic while 
handling cell lines, the level of antibiotics or antimycotic should be tested at the end of the culturing 
process.  
5.1.2 Isolation and purification of primary cells  
5.1.2.1  The biobank shall establish, implement, and maintain procedures for receiv ing and processing 
sources of primary cells.  
NOTE The sources of primary cells include tissue, bone marrow, blood (e.g. mobilized peripheral blood, 
umbilical cord blood), and embryos.  
5.1.2.2  The biobank shall retain additional source materials for confirm ation of origin and, when 
applicable, for histopathological confirmation.  
5.1.3 Primary cultures  
5.1.3.1 Culturing  
For cell culturing, the biobank shall use appropriate nutrients and additives that can optimize the output of 
cell culture in terms of its fi tness for the intended purpose (e.g. reproducibility, stability, or yield).  
5.1.3.2 Characterization  
The biobank shall characterize primary cultures in accordance with their cell types and intended uses. 
The biobank should perform identification and verifi cation for cell cultures when information of 
characteristic markers is available.  
5.1.4 Cell lines  
5.1.4.1 Establishment of cell lines  
The biobank shall record the method and process used for the establishment of cell lines (e.g. by viral 
infection or spon taneous induction by radiation exposure or chemical treatment).  
5.1.4.2 Authentication and characterization   
 The biobank shall authenticate and characterize newly established cell lines to confirm their identities and 
to test for the presence of contaminant s, as suggested as follows:  
a) tests for defining unique attributes;  
b) microbial contamination: visual examination, PCR detection, fluorescence -based DNA detection, 
culture test, enzyme immunoassay, microscopic observation, or an equivalent method;  
c) spe cies verification and authenticity: short tandem repeat (STR) analysis, karyotype analysis, DNA 
barcoding, or an equivalent method.  
5.2 Reception of established cell lines  
5.2.1 General  
5.2.1.1  For the reception of cell lines, the requirements of TZS 2975 , 7.3.2 and A.2 apply. The biobank 
shall establish, implement, and document procedures for the reception of cell lines according to TZS 
2975 , 7.3.2.1.  
5.2.1. 2 Access principles shall be defined in accordance with TZS 2975 , 7.3.1. The biobank should have 
reception requirements, procedures, and forms available, preferably online, to clarify the rights and 
responsibilities of the biobank and the provider.  
5.2.1. 3 Traceability of the cells and its associated data shall be ensured in accordance with TZS 2975 , 
7.5. 
5.2.1 .4 The biobank may receive requests for deposit from an external provider or request deposit from a 
provider. The biobank shall record all deposit requests for future references.  
5.2.2 Review of the deposit request  
5.2.2 .1 The biobank shall review d eposit requests to determine the adequacy of the cell line to be 
deposited, based on the following criteria:  
a) legal considerations or requirements, such as intellectual property issues, the right to distribute and 
dispose, or any contractual obligations;  
b) pre -analytical information (e.g. culture method and condition, preservation method, and storage 
condition);  
c) biosafety level of the cell lines;  
d) pre -defined quality criteria (e.g. free from contamination from foreign organisms, free from hazardous 
material to human health, scientific description including morphology, taxonomic designation, etc.), if 
necessary.  
5.2.2. 2 The biobank shall obtain available characterization information on the cell lines from the provider.  
5.2.2. 3 The biobank should ensur e that a documented agreement or legally binding document is used 
when the cell line is accepted for deposit from the provider. Such agreements describe the rights and 
responsibilities, and immunities of the provider and the biobank with regard to distribu tion and intellectual 
property of the cell line.  
5.2.2.4  The biobank should provide interested parties with access criteria for cell lines at the biobank.  
5.2.3 Decision on the deposit request   
 5.2.3.1  The biobank shall decide whether to accept or to decline the deposit request based on the 
prescribed po licy of the biobank and the result of the review process. The biobank shall document the 
results of the reception reviews.  
5.2.3.2  When the cell line does not conform to the biobank’s criteria, the biobank should notify the deposit 
requestor.  
5.2.4 Transfe r of material and associated data  
5.2.4.1  Once the biobank has decided to accept a cell line for deposit, the biobank may discuss logistical 
issues with the provider to establish a transfer date of the cell line and the associated data, and the 
amount of m aterial to be deposited. The amount of the material to be deposited shall be determined 
according to the nature of the material, the efficacy of preservation, the storage and thawing techniques, 
and the conditions applied on the transfer. Recommended depos it units for the reception of cell lines are 
included in A.1.  
5.2.4.2  The biobank should record the amount of units received from the provider and reserve some units 
for the characterization test. The rest should be stored separately from the existing coll ection until the 
characterization test is completed.  
5.2.4.3  If the cell line is suspected to be either misidentified, cross -contaminated or contaminated by 
microbes, the biobank shall ask the provider or a qualified laboratory to examine the identity of t he cell 
line or to test for the presence of contamination and determine its type during the review process. The 
biobank should confirm if the deposited cell line is previously known to be cross -contaminated or 
misidentified. Biobanks shall be aware of the limit of a detection method at an authentication process.  
NOTE The biobank can check the International Cell Line Authentication Committee (ICLAC) database to 
identify cell lines known to be cross -contaminated . 
5.2.4.4  The biobank shall capture and generate  a structured and standardized report of mammalian cell 
line data.  
5.2.4.5  The biobank shall retain related data on the cell line, including the following, as relevant and 
appropriate:  
a) donor information: donor species, origin, ethnicity, sex, age, colle cted part (e.g. tissue, blood, etc.), and 
disease state, therapy received;  
b) cell line establishment: date of establishment, name and contact information of the originating 
institution, and reference for the establishment;  
c) cell line validation: quality  control testing data (see 5.2.5);  
d) cell line characterization (see 5.2.5);  
e) culture conditions: culture medium, supplement, cryopreservation medium, recommended container for 
storage, and growth condition (e.g. temperature and gas condition);  
f) subcu lture information: passage number or population doubling time, population doubling level, and 
subculture requirements (e.g. split ratio).  
5.2.5 Characterization and authentication of the cell line  
5.2.5.1 The biobank shall perform pertinent characterizatio n tests to verify the viability of the cell line, the 
absence of contaminants, or other characteristics reported by the provider. Such tests can include but are 
not limited to the following:  
a) identity verification: short tandem repeat (STR) analysis for human cell lines or an equivalent method;   
 b) cell morphology: microscopic observation at sparse and confluent conditions;  
c) growth: growth curve and population doubling test or an equivalent method;  
d) microbial contamination: visual examination, PCR dete ction in accordance with TZS 2973 , 
fluorescence -based DNA detection, culture test, enzyme immunoassay, or an equivalent method ; 
NOTE For detection of mycoplasma, mycoplasma colony assay by culturing, PCR, ELISA, and Hoechst 
stain can be used. For other bac teria and fungi, visual examination can be performed at minimum. For virus, 
PCR can be performed . 
e) species verification: isoenzyme analysis, karyotype analysis, DNA barcoding, or an equivalent method;  
f) viability: trypan blue exclusion test, flow cytome try, or an equivalent method.  
g) mutation and copy number variation genotyping (e.g. for tumour cell lines).  
5.2.5. 2 For stem cells, the following additional tests should be performed:  
1) test for differentiation capability;  
2) gene expression signature: q PCR or an equivalent method;  
3) immunophenotyping: fluorescent microscopy or an equivalent method;  
4) test for unique cell surface markers and/or unique cell biofunctions, when applicable.  
5.2.5. 3 If a growth test [see 5.2.5 c)] is performed, the biobank s hall conduct the cell counting in 
accordance with TZS 2974 -1 and TZS 2974 -2 
5.2.5. 4 The biobank shall select and record which characterization tests to perform, based on the types of 
the cell lines.  
5.2.5. 5 The biobank shall perform sufficient characteriza tion tests to confirm that the cell line is authentic 
and free of contaminants (e.g. mycoplasma) and can be used as a valid research tool.  
5.2.5. 6 The biobank shall record the date, the passage number, the test method used, and the results of 
characterizat ion tests. If the test results do not match the characteristic information submitted by the 
provider, the biobank may ask the provider to resubmit the cell line.  
5.2.5. 7 The biobank may decline to accept the cell line if the characteristics do not match th e description 
submitted by the provider or the provider fails to resubmit the cell line. In such cases, the biobank shall 
notify the provider of the decline of acceptance and define how the submitted cell line will be handled, 
either transferring it back t o the provider or disposing it in accordance with applicable requirements and 
policies.  
5.2.6 Assignment of accession number  
When the biobank decides to accept the deposited cell line, the biobank shall assign a unique 
identification number and record the information (see 5.1.1.6 and 5.3.2). When possible, relevant 
information regarding the cell line should be added to an international catalogue.  
5.3 Cell line management  
5.3.1 Planning for cell banking  
The biobank shall establish criteria and methods for pr opagation, preservation, and storage of cell lines in 
accordance with their characteristics, stability, ease of distribution, usage frequency, and difficulty of 
culturing. The biobank should establish short - and long -term plans for storage of cell lines, i n accordance  
 with the current amount of cell lines in the inventory, storage scheme, expected amount of distribution, 
and criteria for characterization and quality control test.  
5.3.2 Accession  
5.3.2.1  The biobank shall determine which data are required fo r accessioning of cell lines (see 5.2.4.5) 
and which data can be considered as supplementary, and collect data accordingly.  
5.3.2. 2 The biobank shall assign a unique identifier in accordance with TZS 2975 , 7.5.1, a) to each cell 
line and record the informa tion at its accession to a database. The following data shall be included in the 
record:  
a) accession date;  
b) accession number;  
c) batch number;  
d) location data;  
e) original number assigned by previous handler(s), if relevant;  
f) previous accession numbe r assigned by the biobank, if relevant;  
g) name and contact informati on of the responsible personnel;  
h) passage number.  
5.3.2. 3 The biobank shall properly tag vials containing cell lines in accordance with TZS 2975 , 7.5.1, a). 
The label should contain the  name of the cell line, accession number, batch number, and other necessary 
data.  
5.3.2. 4 The biobank shall determine whether the cell line and/or related data should be made available for 
distribution. If the cell line is intended for distribution, the bi obank should list the cell line in a catalogue.  
5.3.3 Propagation of cell lines  
5.3.3.1  The biobank shall establish, implement, maintain, and document the relevant procedures for 
culturing cell lines, including necessary preparations of media and consumabl es. The biobank should 
validate or verify the relevant procedures.  
5.3.3. 2 Prior to culturing cell lines, the biobank shall prepare the cell line intended for culturing (e.g. 
thawing frozen cell lines) according to the relevant procedures in order to facil itate high cell survival. The 
biobank should reproduce the original culture conditions of cell lines, as implemented at their 
establishment, since the change in culture conditions can alter the cell behaviour and selection condition. 
The biobank shall docu ment all changes to the procedures.  
5.3.3. 3 The biobank shall record the culture date, identity information of the personnel performing the 
culturing, and the culturing conditions, such as medium composition and percentage, serum type (batch 
number), tempe rature, gaseous phase, and any additional supplement. Personnel responsible for 
culturing shall be aware of and implement measures to prevent various types of contamination.  
5.3.3. 4 While  culturing cell lines, the biobank should frequently inspect their mo rphology and growth and 
perform testing for contamination by mycoplasma, bacteria, mould, or yeast. The biobank shall perform 
frequent tests for identifying common contaminants of cell lines.  
5.3.3. 5 When contamination is detected in a cell line (i.e. micr obial contamination or contamination with 
another cell line), the biobank should check if other batches are also contaminated. The biobank may 
dispose of contaminated cell lines if at least one other batch is uncontaminated. If, however, there is no  
 uncont aminated batch, the biobank can request another deposit from the provider or perform removal of 
contaminants (e.g. with antibiotics) and re -culture the cell line. The biobank shall record and track 
contamination events.  
5.3.3. 6 The biobank should manage th e amount of cell lines in production and the interval between 
productions to ensure that the biobank can consistently provide the requested cell line. In case there is 
evidence in favour of limited passage numbers, the biobank should make a sufficient numb er of batches 
in each production.  
5.3.3. 7 Sub culturing  should occur in accordance with the cell line growth profile.  
5.3.4 Preservation and storage  
5.3.4.1  The requirements of TZS 2975 , 7.6, 7.7, A.4 and A.6 apply.  
5.3.4. 2 The biobank shall establish, imp lement, maintain, and document cryopreservation method and 
storage condition for cell lines, taking into account of characteristics and viability of each cell line. The 
biobank should validate or verify cryopreservation and storage methods. The biobank sho uld perform 
quality control (QC) tests on each batch of the preserved cell stocks.  
5.3.4. 3 The biobank should implement seed stock approaches that allow the separate storage of cell lines 
into a seed stock for long -term storage and a distribution stock for  short -term storage and frequent uses. 
The biobank should set the number of vials produced for each stock and minimum number of vials 
required to be in stocks prior to the replenishment of vials (see Table 1). The biobank should store an 
aliquot of each ba tch or each stock.  
Table 1 — Example of cell line storage condition  
Purpose  Storage condition  Minimum cell number 
per vial  Minimum number of 
vials  
Seed stock  Vapour -phase of liquid 
nitrogen(recommended)  
(≤-130 °C)  1 × 106 cells/ml  
(each vial)  12 
Distribu tion stock  20 
 
5.3.5 Inventory management  
5.3.5.1  Requirements  of TZS 2975 , 7.5.1 c) apply.  
5.3.5. 2 The biobank should implement a computer -based system for managing its inventory. The biobank 
should immediately update the database when the biobank rece ives cell lines, transports them within the 
biobank, or distributes them to others outside the biobank. A back -up should be available.  
5.3.5. 3 The biobank should verify the cell line inventory at planned intervals by a defined procedure. The 
biobank shall periodically inspect the storage and label conditions by eye or by using a rack scanner. In 
case of any discrepancy between the record and actual status, the biobank shall examine the cause of 
such an inconsistency, document it, and modify the record to re flect the actual status.  
5.3.6 Disposal management  
5.3.6.1  The requirements of TZS 2975 , 4.1.8, 7.1.1, 7.5.3 and A.7 apply.  
5.3.6. 2 The biobank shall establish, implement, and document criteria for disposing cell lines, such as for 
the following conditions :  
 a) use of cell lines is no longer permitted due to legally binding agreements;  
b) withdrawal of consent, where required;  
c) cell lines are altered from their original states during processing;  
d) cell lines are contaminated and no longer considered usefu l; 
e) storage of cell lines is no longer feasible or necessary.  
5.3.6. 3 The biobank shall store cell lines identified for disposal at a separate space or, at least, in a 
separate container until their collection by a specialized waste service.  
5.4 Distribu tion 
5.4.1 General  
The requirements of TZS 2975 , 7.3.3 apply. Recommended units for the distribution of cell lines are 
included in A.2.  
5.4.2 Acceptance of order request  
The biobank should have distribution requirements, procedures, and forms available, pr eferably online, to 
clarify the rights and responsibilities of the biobank and the recipient/user.  
The biobank should accept distribution requests of qualified recipient/users and shall record the following 
information:  
a) request number and date;  
b) conta ct information of the requested user and, if applicable, the institutional official;  
NOTE Contact information includes but is not limited to name, affiliation, address, phone number, 
and email address . 
c) the requested cell line, including name, amount, an d volume.  
5.4.3 Distribution review  
The biobank shall establish and publish criteria for requestors/users to access the cell lines. The biobank 
shall check the eligibility of the requestor/user based on these criteria. The biobank shall record all order 
requests and notify the decision to the requested user.  
5.4.4 Transport  
5.4.4.1 General  
The requirements of TZS 2975 , 7.4 apply. The biobank shall document the transport temperature.  
5.4.4.2 Preparation for shipment  
The biobank should prepare for shipment of  the requested cell line after confirming the inventory record 
and results of quality control tests. In case the cell line is shipped as cultures for local distribution, the 
biobank shall ensure viability of a substantial proportion of the cell line prior to the shipment.  
5.4.4.3 Packaging  
5.4.4.3 .1 The biobank shall package the cell line in a container that is resistant to damage and leakage 
and with relevant safety labels included both within and outside of the package.   
 5.4.4.3. 2 The biobank shall impleme nt procedures to maintain the frozen status of the cell line until the 
recipient/user receives it. The biobank shall use a type and amount of coolant appropriate to the type of 
cell line  and needs of the recipient/user. Personnel shall use personal protect ive equipment when 
handling cell lines and coolants.  
5.4.4.3. 3 If the cell line is sent in a culture flask for local distribution, personnel shall fill up the culture flask 
with culture medium and tightly seal the lid with tape, or similar material to prev ent leakage. Personnel 
should further wrap the flask with absorbent cotton and put it in a zipped bag and an outer box 
sequentially to prevent leakage and maintain the required temperature.  
5.4.4.4 Shipment  
The biobank should send out the requested cell li ne in a timely manner and notify the recipient/user in 
case the biobank cannot provide the requested cell line or needs extra time for culturing. If a microbial or 
cross contamination is detected in the distributed cell line after shipment, the biobank sha ll notify the 
recipient/user of the cell line.  
5.5 Quality control, validation and verification  
5.5.1 Quality control  
5.5.1.1 The biobank shall conduct quality control for the quality of cell lines in accordance with TZS 2975 , 
7.8. 
5.5.1. 2 The biobank shal l establish criteria for cell lines requiring quality control tests and plan to execute 
the relevant tests. The biobank should perform quality control tests on samples from each stock.  
5.5.1. 3 QC should be done prior to freezing the cells.  
5.5.1. 4 The biob ank should periodically monitor cell lines for mycoplasma contamination. See 5.2.5 for 
the list of quality control tests for cell lines.  
5.5.1. 5 Whenever the cell line has to be assessed for quality control tests, the biobank should record the 
relevant dat a, such as accession numbers of the cell lines, storage locations, executed tests, and their 
results and uncertainties.  
5.5.1. 6 The biobank should establish alternative procedures for cell lines that cannot be tested for quality 
control or for which additi onal tests are required.  
5.5.1. 7 The biobank shall perform evidence -based stability studies for long - and short -term storage of cell 
lines.  
5.5.1. 8 In case the test result reveals that the examined cell line is altered from its original state, the 
biobank should perform additional tests on the cell line from the identical batch. If additional tests reveal 
the same sign of alteration, the biobank shall handle the altered cell lines according to its policy and 
record the occurrence.  
5.5.2 Validation and verif ication  
5.5.2.1  The biobank shall validate and/or verify their methods in accordance with TZS 2975 , 7.9.  
5.5.2.2  The biobank shall document and retain the validation/verification results, as well as the 
information about the procedures used, and descriptio ns of the appropriateness of the method of use.  
5.5.2.3  The biobank should supervise the validation and process control schedules.  
  
 6 Performance evaluation and improvement  
6.1 The biobank shall perform an internal evaluation at planned intervals to improv e efficiency of its 
management system and services regarding cell lines.  
6.2 The biobank shall evaluate the performance based on its compliance to this document and guidelines 
established by the biobank.  
6.3 The biobank shall review outcomes of t he evaluation and establish improvement schemes for the 
biobank.  
 
 
 
Annex A  
(informative)  
Recommended units for biobanking of cell lines  
A.1 Deposit unit for reception of cell lines  
The recommended deposit unit is as follows:  
a) Frozen: a minimum of 3 vial s to 5 vials from the same batch/unit; each containing minimum of 1 × 106 
cells/mL; maximum of 1 × 107 cells;  
b) Culture: 3 culture flasks (T75 or T25) from the same batch/unit; each containing minimum of 1 × 106 
cells/ml; maximum of 1 × 107 cells.  
A.2 Dis tribution unit of cell lines  
The recommended distribution unit is as follows:  
a) Frozen: 1 vial; each containing minimum of 1 × 106 cells/mL; maximum of 1 × 107 cells;  
b) Culture: 1 culture flask (T25); each containing minimum of 1 × 106 cells/ml; maximum of 1 × 107 cells.