ZenBio, Inc.

Human Adult Stem Cell Manual
Differentiation protocols for humanadipose-derived adult stem cells

INSTRUCTION MANUALZBM0015.04

SHIPPING CONDITIONSHuman Adult Stem Cells (ASC)
Orders are delivered via Federal Express courier. All US and Canada orders are shipped via
Federal Express Priority service and are usually received the next day. International orders areusually received in 2-4 days. Please inquire if alternate couriers are needed.
Must be processed immediately upon shipment receipt.

STORAGE CONDITIONS

 Media: Short Term 4°C
6 months-20°C
 Cryopreserved cells: Vials of frozen preadipocytes are to be stored in vapor phase
nitrogen (-150C to -190C).

All Zen-Bio Inc products are for research use only. Not approved for human or veterinary use orfor use in diagnostic or clinical procedures.

ORDERING INFORMATION AND TECHNICAL SERVICES

ZenBio, Inc.
3200 East NC-54 Suite 100
PO Box 13888
Research Triangle Park, NC 27709
U.S.A.
Telephone(919) 547-0692
Facsimile (FAX)(919) 547-0693
Toll free (continental US only)
1-866-ADIPOSE 1-(866)-234-7673
Electronic mail (e-mail)information@zenbio.com
World Wide Web
http://www.zenbio.com

THIS MANUAL IS SUITABLE FOR USE WITH THE FOLLOWING PRODUCTS:
ASC-F, ASC-F-SL

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CONTENTSPAGE #

Introduction

3

Materials Provided for Each Catalog Item

3

Media Compositions

4

Plating and Expansion of Cryopreserved Adult Stem Cells

5

Adipogenesis Protocol

6

Osteogenesis Protocol

8

Chondrogenesis Protocol

10

Troubleshooting

11

Frequently Asked Questions

12

Pathogen testing

13

LIMITED PRODUCT WARRANTYThis warranty limits our liability to replacement of this product. No other warranties ofany kind, expressed or implied, including without limitation implied warranties ofmerchantability or fitness for a particular purpose, are provided by Zen-Bio, Inc. Zen-Bio, Inc.
shall have no liability for any direct, indirect, consequential, or incidental damages arising outof the use, the results of use, or the inability to use this product.
Zen-Bio, Inc warrants the performance of cells only if Zen-Bio media are used and therecommended protocols are followed. Cryopreserved cells are assured to be viable whenthawed according to Zen-Bio protocols and using the recommended cultureware.
Contact ZenBio, Inc. within no more than 24 hours after receipt of products for all claims
regarding shipment damage, incorrect ordering or other delivery issues. Delivery claims
received after 7 days of receipt of products are not subject to replacement or refund.
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PRECAUTIONSThis product is for research use only. It is not intended for human, veterinary, or in vitro
diagnostic use. Proper precautions and biological containment should be taken when handlingcells of human origin, due to their potential biohazardous nature. Always wear gloves andwork behind a protective screen when handling primary human cells. All media,
supplements, and tissue cultureware used in this protocol should be sterile.
Human adult stem cell viability depends greatly on the use of suitable media, reagents,and sterile plastic wear. If these parameters are not carefully observed, limited differentiation
may occur and cell growth may be slow.

INTRODUCTION
The adipose derived adult stem cells are isolated from subcutaneous adipose tissue of
healthy non-diabetic donors between 18 and 60 years old undergoing elective surgery. The
cells are isolated by centrifugal force after collagenase treatment. Adult stem cells can bedifferentiated into various lineages using Zen-Bio media formulations and protocols.

This

instruction manual describes procedures to induce human adipose derived adult stem cells(ASC) to differentiate into 1) mature adipocytes, 2) osteoblasts, and 3) chondrocytes. The
process of differentiating human adipose-tissue derived adult stem cells to adipocytes has
been patent protected by Zen-Bio under US patent number 6153432.

MATERIALS PROVIDED FOR EACH CATALOG ITEM
Cryopreserved Human Adipose-Derived Adult Stem Cells(catalog # ASC-F, ASC-F-SL)
Frozen vial containing 1.0 viable adult stem cells (store in vapor phase liquid nitrogen uponreceipt)oEffective February 1, 2016, we will no longer provide 50ml support medium with eachcryopreserved vial ordered. Please order media according to your needs.

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MEDIA COMPOSTIONSPreadipocyte Medium
cat # PM-1
DMEM / Ham’s F-12 (1:1, v/v)
HEPES pH 7.4
Fetal Bovine Serum (FBS)
Penicillin
Streptomycin
Amphotericin B

Adipocyte Basal Medium
Cat# BM-1
DMEM / Ham’s F-12 (1:1, v/v)
HEPES pH 7.4
BiotinPantothenate

Adipocyte Differentiation
Mediumcat # DM-2

Adipocyte MaintenanceMediumcat # AM-1

DMEM / Ham’s F-12 (1:1, v/v)
HEPES pH 7.4
Fetal Bovine Serum (FBS)DMEM / Ham’s F-12 (1:1, v/v)
HEPES pH 7.4
Fetal Bovine Serum (FBS)

BiotinPantothenate
Human insulin
Dexamethasone
Isobutylmethylxanthine
PPAR agonist
Penicillin
Streptomycin
Amphotericin B

BiotinPantothenate
Human insulin
Dexamethasone
Penicillin
Streptomycin
Amphotericin B

Osteoblast Differentiation
MediumCat# OB-1

DMEM / Ham’s F-12 (1:1, v/v)
Fetal Bovine Serum (FBS)-glycerophosphate
Ascorbate-2-phosphate
Dexamethasone
1,25 (OH)2 Vitamin D3
Penicillin
Streptomycin
Amphotericin B

Chondrocyte Differentiation
MediumCat# CM-1

DMEM-high glucose
Fetal Bovine Serum (FBS)
Transforming growth factor 1(TGF-1)
Ascorbate-2-phosphate
Dexamethasone
Insulin-transferrin-selenium plus (ITS+)
Penicillin
Streptomycin
Amphotericin B

NOTE:
All media except Chondrocyte Medium contains 3.15g/L (17.5mmol/L) D-glucose.
Chondrocyte Differentiation Medium contains 4.5g/L (25.0mmol/L) D-glucose
All media are also available as phenol red free and/or without serum. Please inquire forcustom media requests.

MEDIA EXPIRATION DATES:
If placed at 4C upon arrival, the media is stable until the expiration date on the bottle
label.
If stored at -20C upon arrival, the media is stable for 6 months. Add fresh antibiotics
when you are ready to use. The media will expire 30 days after the thaw date.
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PLATING AND EXPANSION PROCEDURES
Cryopreserved Human Adult Stem Cells

Please note: Primary cells can be very sensitive to brands of cultureware. Zen-Bio does not currently
recommend the use of Corning Falcon or Sarstedt brand plates or flasks. Our scientists are using
Nunc, Corning Costar, or Greiner Bio-One Cellstar tissue culture treated plates and flasks. Pleasecontact us if you have any questions.
1. Remove cells from liquid nitrogen and place immediately into a 37C water bath with agitation.

Be careful not to submerge the cap of the vial into water. Do not leave the vials in water bathafter most of the content has thawed. Rinse the vials with 70% ethanol before taking them tothe culture hood.
2. Upon the thawing, add the cells to a sterile conical bottom centrifuge tube, containing 10 ml of

Growth Medium (Preadipocyte Medium (PM-1).
3. Centrifuge at 280 x g, 20C, 5 minutes. Aspirate the medium and resuspend cells in a volume of

PM-1 appropriate for counting the cells. Count using a hemacytometer.
4. Place approximately 6.7 X 105 cells in T-75 culture flasks using PM-1.
5. Incubate cells until they are 85-90% confluent (in about 4-5 days). Do not let the cells become

100% confluent (see Figure 1-A, page 8 for picture of 100% confluent cells). Cells will need to
be fed every other day with PM-1.
6. Aspirate medium and wash adult stem cells 4-5 times using sterile Phosphate Buffered Saline(PBS) to remove all traces of serum (until there is no foaming of the medium). Remove the PBS
and release the cells from the flask bottom by adding 2 mL/T-75 flask (or 6 ml/T-225 flask) of
0.25% trypsin/ 2.21mM EDTA solution. Allow cells to trypsinize for 5 minutes at 37C. Tap theflask gently to loosen the cells.
7. Neutralize the trypsin using 7 ml PM-1 per T-75 flask (or 21 ml per T-225 flask). Check the flask

under a microscope to ensure all cells are free of the flask bottom.
8. Count the cells and plate in desired format. Ensure cells are evenly suspended when platinglarge numbers of plates or flasks. Do not agitate plates and flasks after plating. Place in ahumidified incubator at 37C and 5% CO2, making sure the surface is level for even cell
distribution.
9. OPTIONAL – Cryopreserve stem cells after counting. Centrifuge at 280 x g, 20C, 5 minutes.

Suspend in cold cryopreservation medium (Cat# FM-1-100) at a concentration of 1X106 cells/ml.
Do not exceed a 6:1 ratio of cells (per million): volume cryopreservation medium (per ml).
Remember to account for the volume of the cell pellet before adding the volume of
cryopreservation medium necessary for cell suspension. If using a controlled-rate freezer:
Freeze by reducing the temperature 1°C per minute until the temperature reaches -80° C. If
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using a cell cryopreservation container, prepare according to the manufacturer’s instructions.
For best results we recommend transferring the vials to the vapor phase of a liquid nitrogenstorage facility as soon as possible after the cells have reached -80°C.

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ADIPOGENESIS PROCEDURE
Differentiation of Adult Stem Cells into Adipocytes
Please note: Primary cells can be very sensitive to brands of cultureware. Zen-Bio does not
currently recommend the use of Falcon or Sarstedt brand plates or flasks. Our scientists areusing Nunc, Costar/Corning, or Greiner bio-one CellStar tissue culture treated plates and flasks.
1. Remove cells from liquid nitrogen and place immediately into a 37 C water bath and agitate
while in bath. Be careful not to submerge the cap of the vial into water. Do not leave the vials
in water bath after most of the content has thawed. Rinse the vials with 70% ethanol before
taking them to the culture hood.
2. Upon thawing, transfer the cells to a sterile conical bottom centrifuge tube containing 10 ml ofPreadipocyte Medium (cat # PM-1). Centrifuge: 1,200 rpm (282 X g) / 20C / 5 minutes.
Aspirate the supernatant. TAKE CARE TO NOT ASPIRATE ANY OF THE CELL PELLET.
3. The cell vial contains a minimum of 1.0 or 2.0 x 106 viable cells; however, we recommend
performing a cell count to determine a more exact number of cells. Resuspend the cell pellet in
2 ml Preadipocyte medium; dilute an aliquot in 0.4% trypan blue solution. We suggest
withdrawing an aliquot of 50 l of cells and mixing with 100 l of the trypan blue solution,
resulting in a dilution factor of 3. Count live (unstained) cells on a hemacytometer.
4. Plate approximately 40,625 cells / cm2 using the media volumes from the table below. Refer tothe manufacturer’s specifications for the specific cultureware brand you are using.
FORMAT

VOLUMEPER WELL

96 well plate
48 well plate
24 well plate
12 well plate
6 well plate
10 cm dishT-75 flask
T25 flask

150 l500 l
1 ml
2 ml
3 ml
15 ml
20 ml
7 ml

TOTAL
VOLUME PERFORMAT*
14.4 ml24.0 ml24.0 ml24.0 ml18.0 ml
15.0 ml
20.0 ml
7.0 ml*We recommend preparing slightly larger volumes to allow for loss due to foam and pipet error.
5. Plate cells in desired format and place in a humidified 37oC incubator with 5% CO2. Do notagitate the plate, as cells will not plate evenly.
6. Twenty-four hours after plating, check the plates for confluence. If they are not completely
confluent, leave for and additional 24 hours maximum before inducing differentiation. If the cells
are not confluent after 48 hours, DO NOT INDUCE DIFFERENTIATION (differentiation will bepoor). Contact Zen-Bio immediately.

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7. To start the process, aspirate the entire volume of Preadipocyte Medium from all wells. Add theappropriate volume of Adipocyte Differentiation Medium (catalog # DM-2) to the wells (seeTable 1. Feeding Volumes). Incubate plate for 7 days at 370C and 5% CO2.
8. After 7 days, cells should be fed by removing some of the media and replacing with fresh
Adipocyte Medium (catalog # AM-1) (See Table 1. Feeding Volumes). Caution: Do not dry thewells. Add new medium gently. If using an automatic feeder, set the slowest flow ratepossible.
9. Two (2) weeks after the initiation of differentiation, cells should appear rounded with large lipid
droplets apparent in the cytoplasm (see Figure 1-C). Cells are now considered matureadipocytes and are suitable for most assays.

Table 1. Adipogenesis: Feeding Volumes
FormatPlatingChange PM-1 to DM-2

Change DM-2 to AM-1

Change AM-1 to AM-1

IN

OUT

IN

OUT

IN

OUT

IN

96 well plate

150 l/well150 l/well150 l/ well

90 l/well120 l/well

90 l/well

90 l/well

48 well plate

500 l/well500 l/well500 l/well300 l/well400 l/well300 l/well300 l/well

24 well plate

1.0 ml/well

1.0 ml/well

1.0 ml/well

0.6 ml/well

0.8 ml/well

0.6 ml/well

0.6 ml/well

12 well plate

2.0 ml/well

2.0 ml/well

2.0 ml/well

1.2 ml/well

1.6 ml/well

1.2 ml/well

1.2 ml/well

6 well plate

3.0 ml/well

3.0 ml/well

3.0 ml/well

1.8 ml/well

2.4 ml/well

1.8 ml/well

1.8 ml/well

T-75 flask

20 ml/flask

20 ml/flask

20 ml/flask

12 ml/flask

16 ml/flask

12 ml/flask

12 ml/flask

T-25 flask

7 ml/flask

7 ml/flask

7 ml/flask

4.2 ml/flask

5.6 ml/flask

4.2 ml/flask

4.2 ml/flask

A. 100% Confluent
Adult stem cells

B. 1-week-old adipocytes(1 wk post-differentiation)

C. 2-week-old adipocytes(2 wks post-differentiation)

PREADIPOCYTE MATURE ADIPOCYTE

nucleusnucleus

Lipid droplets

Figure 1: Photographs of 100% confluent Adult stem cells (ASC) (A), 1-week-old (post-differentiation) cultured
adipocytes (B) and mature (2 weeks post-differentiation) cultured Adipocytes (C). These are unstained photographsof human adult stem cell morphology (20X). The cells should appear comparable in appearance to these pictures.

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OSTEOGENESIS PROCEDURE

Differentiation of Adult Stem Cells into Osteoblasts
NOTE: We recommend using Nunc, Costar/Corning, or Greiner bio-one CellStar tissue culture treatedplates and flasks.
1. Remove cells from liquid nitrogen and place immediately into a 37 C water bath and agitate
while in bath. Be careful not to submerge the cap of the vial into water. Do not leave the vials
in water bath after most of the content has thawed. Rinse the vials with 70% ethanol before
taking them to the culture hood.
2. Upon thawing, transfer the cells to a sterile conical bottom centrifuge tube containing 10 ml ofPreadipocyte Medium (cat # PM-1). Centrifuge: 1,200 rpm (282 X g) / 20C / 5 minutes.
Aspirate the supernatant. TAKE CARE TO NOT ASPIRATE ANY OF THE CELL PELLET.
3. The cell vial contains a minimum of 1.0 or 2.0 x 106 viable cells; however, we recommend
performing a cell count to determine a more exact number of cells. Resuspend the cell pellet in
2 ml Preadipocyte medium; dilute an aliquot in 0.4% trypan blue solution. We suggest
withdrawing an aliquot of 50 l of cells and mixing with 100 l of the trypan blue solution,
resulting in a dilution factor of 3. Count live (unstained) cells on a hemacytometer.
4. Plate approximately 30,000 cells / cm2 using the media volumes from the table below. Refer tothe manufacturer’s specifications for the specific cultureware brand you are using.
FORMAT

VOLUMEPER WELLTOTAL VOLUME PERFORMAT

NUMBER CELLS PERFORMAT

96 well plate

150 l15 ml /plate

0.93 X 106 cells/15 ml

48 well plate

500 l
1 ml
2 ml
3 ml
15 ml
20 ml
7 ml

24ml/plate1.25 X 106 cells/25 ml

25ml/plate
25ml/plate
18 ml/plate
15ml/dish
20 ml/flask
7 ml/ flask

1.44X 106 cells/26ml
1.5 X 106 cells/26ml
1.8 X 106 cells/20ml
2.25 X 106 cells/15 ml
2.25 X 106 cells/20ml
0.75 X 106 cells/7ml

24 well plate
12 well plate
6 well plate
10 cm dishT-75 flask
T25 flask*We recommend preparing slightly larger volumes to allow for loss due to foam and pipet error.
5. Plate cells in desired format and place in a humidified 37oC incubator with 5% CO2. Do notagitate the plate, as cells will not plate evenly.
6. Twenty-four hours after plating, aspirate the entire volume of Preadipocyte Medium from allwells. Add the appropriate volume of Osteoblast Differentiation Medium (catalog # OB-1) to thewells (see Table 3. Osteogenesis Feeding Volumes).
7. Incubate cells at 370C and 5% CO2.
8. Feed cells every 3 days with Osteoblast Differentiation Medium (catalog # OB-1).

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Table 3. Osteogenesis Feeding Volumes
FormatVolume per well
96 well plate
150 l/well
48 well plate
24 well plate
12 well plate
6 well plate
T-75 flask
T-25 flask

300 l/well
1.0 ml/well
2.0 ml/well
3.0 ml/well
12 ml/flask
7 ml/flask

Figure 2. Osteoblasts, (20X)
Alizarin red staining of 14 day old cells shows significant mineralization.

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CHONDROGENESIS PROCEDURE

Differentiation of Adult Stem Cells into Chondrocytes
NOTE: We recommend using Nunc, Costar/Corning, or Greiner bio-one CellStar tissue culture treatedplates and flasks.
NOTE: You will need to prepare
1. Remove cells from liquid nitrogen and place immediately into a 37 C water bath and agitate
while in bath. Be careful not to submerge the cap of the vial into water. Do not leave the vials
in water bath after most of the content has thawed. Rinse the vials with 70% ethanol before
taking them to the culture hood.
2. Upon thawing, transfer the cells to a sterile conical bottom centrifuge tube containing 10 ml ofPreadipocyte Medium (cat # PM-1). Centrifuge: 1,200 rpm (282 X g) / 20C / 5 minutes.
Aspirate the supernatant. TAKE CARE TO NOT ASPIRATE ANY OF THE CELL PELLET.
3. The cell vial contains a minimum of 1.0 or 2.0 x 106 viable cells; however, we recommend
performing a cell count to determine a more exact number of cells. Resuspend the cell pellet in
2 ml Preadipocyte medium (PM-1), dilute an aliquot in 0.4% trypan blue solution. We suggest
withdrawing an aliquot of 50 l of cells and mixing with 100 l of the trypan blue solution,
resulting in a dilution factor of 3. Count live (unstained) cells on a hemacytometer.
4. Remove the supernatant and resuspend the cell pellet in 1.2 % alginate solution (made in
150mM NaCl) at 4 x 106 cells/ml. Pipette up and down without creating bubbles to mix
thoroughly.
5. Draw the cell-seeded alginate suspension into a sterile 10 cc syringe using a 22 gauge needle.
6. Add 3 ml of CaCl2 (102 mM) into one well of a 6 well culture plate.
7. Carefully and slowly dispense equal-sized droplets of the cell-seeded alginate solution into the
CaCl2 solution (dispense10-30 beads per well taking care to avoid clumping the beads). Curethe beads in the CaCl2 solution for 10 minutes at room temperature.
8. Using a glass pipette, aspirate the CaCl2 solution off the beads. Be careful not to aspirate thebeads with the solution.
9. Wash the cell-seeded alginate beads three times with NaCl (150 mM) and then one more timewith DMEM-HG.
10. Add 3 ml of the Chondrogenic Differentiation Medium (cat# CM-1).
11. Incubate at 37°C, 5.0 % CO2, and 95% relative humidity for the duration of the experiment.
12. Change media every three days.
Figure 3. Differentiated cells stainedfor collagen production.

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TROUBLESHOOTING GUIDE
Observation

Possible causes

Adult stem cells do notdifferentiate

1. Cells have been passaged
too many times

1. Use cells of a lower passage
number

2. Differentiation conditions not
optimal

2. Use our defined differentiation
medium.

3. Cells were plated at a low
density

3. Use the cell density recommended
in our manual

4. Cultureware used not
optimal for human primaryadipocytes

4. Zen-Bio does not recommend theuse of Falcon or Sarstedt brandcultureware for all Adipogenesis
cell culture applications

5. Differences in cultureware
brand surface area may
affect plating density inunknown
1. Cells have been passaged
too many times
2. Cells expanded too high
3. Cultureware used not
optimal for human primary
adult stem cells
1. Medium in outside wells
evaporated

5. Verify the surface area for thecultureware brand you are using.

Adult stem cells do not
grow

Edge effects

Adipogenesis:
Adipocytes appear
uneven in each well

1. Medium was completely
removed during feeding
2. Fresh medium was added
too quickly
3. Cells placed on unevensurface in the incubator

Suggestions

1. Use cells of a lower passage
number
2. Do not exceed 1:6 expansion ratio
3. Use only Costar, Nunc or Greiner
cultureware
1. Ensure a saturated humidity in the
incubator. Make sure multiple
plates are stacked no more than 3plates high.
1. Make sure to follow instructions
listed in Table 1 Feeding Volumes
2. Add media slowly to each well.
Position the pipet tips halfway
down, pressing on the side of thewells and slowly release themedium.
3. Place cultureware are on a levelsurface in the incubator to ensure
cells attach evenly.

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FREQUENTLY ASKED QUESTIONS
1Can I pass the cells?

Adult Stem Cells can be trypsinized and re-plated.
They grow slower with each passage and undergo
adipogenesis poorly after passage 4.

2

How fast do the cells replicate?

All cells are shipped at Passage 2-3.
The average doubling time is 48-84 hours. However,keep in mind that the replication rate varies slightly

3

Should antibiotics be included in
the medium?

from donor to donor.
Yes. Antibiotics and anti-fungal agents are always
recommended since the cells are primary cells. All ZenBio media contain antibiotics and/or anti-fungal agents4

Where are the cells obtained?

except Adipocyte Basal Medium (cat# BM-1).
The adult stem cells are isolated from humansubcutaneous adipose tissue from consented adult5

Do you test for pathogens?
Which ones?

donors undergoing elective surgeries.
Yes. Samples from each donor are tested via PCR to
confirm non-reactivity for HIV-1, HIV-2, hepatitis B andhepatitis C. However, since we cannot test all
pathogens, please treat the culture as a potentially67

How do I obtain RNA from thecells? How much RNA can I
expect?

What donor information do I
receive?

8What is the formulation of ZenBio’s serum-free media?

infectious agent.
Use RNA Tri-reagent (Molecular Products), RNeasy kit(Qiagen) or other column based kit. You can expect
approximately 20 g total RNA from a 10 cm dish ofundifferentiated adult stem cells.
The donor’s age, gender, and BMI are provided in thecertificate of analysis that accompanies each lot ofcells
Zen-Bio’s serum-free media are not enhanced tosupplement the absence of serum. These media are
available for assay procedures where cells are rested
from serum. Do not differentiate adult stem cells into

9What quality control testing is
performed on the cells?

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adipocytes using medium without serum.
We do confirm the presence of several cell surface
markers indicative of stem cells via flow cytometry.
The adult stem cells stain >99% positive for CD105
and CD44; negative for CD31 and CD45.
Our quality control :-for adipocytes is lipid staining, total triglyceride
content, and functional lipolysis;
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-for osteoblasts it is measurement of degree ofmineralization as assessed by Alizarin Red staining-for chondrocytes it is collagen staining
10

What is the concentration ofingredients in your media?

11

Can the adult stem cells be
differentiated into any
additional cell types?

We do not disclose the concentrations of thecomponents of our media.
- We are happy to prepare custom media to yourspecifications.
Other researchers have differentiated adult stem cells
into smooth muscle, neuronal, and hepatocyte
lineages. See selected references below.
-

 Mizuno, et al. Myogenic differentiation by humanprocessed lipoaspirate cells. Plastic andReconstructive Surgery (2002) 109:199-209.
 Seo, et al. Differentiation of human adipose stromalcells into hepatic lineage in vitro and in vivo.
Biochem Biophys. Res. Comm. (2005) 328: 258264.
 Rodríguez, RV, et al. Clonogenic multipotent stem
cells in human adipose tissue differentiate intofunctional smooth muscle cells. Proc Natl Acad Sci
USA. (2006), 103(32): 12167–12172.
 Wouter, JFM et al. Effect of tissue-harvesting site onyield of stem cells derived from adipose tissue:
implications for cell-based therapies. Cell TissueRes. (2008) 332(3): 415–426.

PATHOGEN TESTING
Samples from each donor are tested via PCR to confirm non-reactivity for HIV-1, HIV-2, hepatitis B
and hepatitis C. However, no known test can offer complete assurance that the cells are pathogen free.
Our products are tested and are free from mycoplasma contamination. Proper precautions andbiological containment should be taken when handling cells of human origin, due to their potential
biohazardous nature. All human based products should be handled at a BSL-2 (Biosafety Level 2) or
higher. Always wear gloves and work behind a protective screen when handling primary human cells.

REFERENCES
Lists of articles using ZenBio, Inc human adult stem cells may be found at our website(http://www.zenbio.com).
株式会社ケー・エー・シー
試薬事業部〒661-0978

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