INSTRUCTIONSFOR USECONTENTS
1. Introduction
2. Safety information
3. Product storage instructions
4. GrowDase is used for the enzymatic degradation of GrowDex
5. Procedure for removing GrowDex by enzymatic degradation
6. Example experimental procedure
7. Dilution table
8. Ordering information
9. Contact information

This product is for research use only and should not be used for diagnostic or therapeutic purposes.

1. INTRODUCTION
GrowDase™ is a purified mixture of cellulase enzymes(10 mg/ml) that has been specifically developed tobreakdown the nanofibrillar cellulose present in UPM’sGrowDex® to soluble glucose. The process is simple,
enzyme is mixed with the sample and incubated at37°C until the GrowDex has been reduced to non-toxicsugars. Enzymatic removal allows cells to be recoveredfrom the matrix efficiently, retain their 3D structure, e.g.
spheroid or organoid in-tact and does not exhibit any
adverse effect on cell viability or functionality. The amountof GrowDase required for cell recovery is dependent
on the amount of GrowDex (cellulose) present in thesample. One GrowDase vial (2.5 ml) can degrade
5 ml of GrowDex. Recovery of cells, organoids or
spheroids for downstream processing once a live cellassay has been concluded can be advantageous formany cell-based studies. Additional data relating
to gene or protein expression can be generated orcells used for further experimental work such as inmulti-dose studies.
GrowDase is for research use only and should not beused for diagnostic or therapeutic purposes.

2. SAFETY INFORMATION
GrowDase consists of a cellulase enzyme mixture inaqueous sodium acetate buffer solution, pH 5. The
product is sterile and intended for the degradation ofGrowDex. GrowDase is for research use only, not for
diagnostic or therapeutic use.
In accordance with current regulations (1272/2008
CLP), this substance has been classified as nonhazardous. The product contains up to 10 mg/ml
enzymes and has been sterile filtered.
The product should be handled in accordance with goodlaboratory practices (GLP). Use protective gloves andclothes to avoid skin exposure. If exposed wash theskin with water. Use protective laboratory eye wear
to avoid contact with the eyes.
Hazard Statements:
EUH208 Contains enzymes. May produce an allergic
reaction.

Description of first aid measures:• Inhalation: Move to fresh air. Seek medical attention
if symptoms appear.
• Skin contact: Wash with water. Seek medical attention
if irritation occurs.
• Eye contact: Rinse with plenty of water for several
minutes. Seek medical attention if irritation occurs.
• Ingestion: Rinse mouth with plenty of water. If large
quantities of the product are ingested endeavour tovomit. Seek medical attention if symptoms appear.
Most important symptoms and effects, bothacute and delayed• Inhalation: May produce an allergic reaction.
• Ingestion: No symptoms or effects known.
• Skin contact: No symptoms or effects known.
• Eye contact: No symptoms or effects known.

NOTE: For further information refer to the GrowDase Material Safety Data Sheet.

3. PRODUCT STORAGE INSTRUCTIONS
The product has a shelf life of 12 months from date ofmanufacture and should be stored at 4-22°C (39–72°F)
and protected from light for optimum performance.

If the product has been diluted, e.g. with culture media,
then it should be used immediately. Any leftover dilutedenzyme should be discarded.

Once opened it is recommended that the product isstored at 4-8°C (39-46°F) for a maximum of 3 months.

4. GROWDASE IS USED FOR THE ENZYMATIC
DEGRADATION OF GROWDEXGrowDase is a purified mixture of cellulase enzymes
that has been specifically developed to reduce thenanocellulose fibrils present in GrowDex to soluble
glucose. The process is simple, the enzyme is mixed withthe sample and incubated at 37°C until the GrowDex
has been fully reduced to glucose. Enzymatic removal
allows cells to be recovered from the matrix efficiently,
retain their 3D structure, e.g. spheroid or organoid,
intact and without adverse effects on cell functionality.
The amount of GrowDase required for cell recovery

Cells cultured
in GrowDex

is dependent on the amount of GrowDex hydrogel(cellulose) present in the sample.
It is recommended that GrowDase is used at a working
concentration of 300 µg/mg of GrowDex. An equalvolume of working concentration GrowDase toGrowDex/cell matrix volume present in the sample
should be used, i.e. if 100 µl of GrowDex/cell matrix
is present in the sample then 100 µl of GrowDase (300µg/mg) should be added to that sample well.

GrowDaseadded, sample
incubated at 37°C

GrowDex degradedto glucose, cells
are in solution

5. PROCEDURE FOR REMOVING GROWDEX HYDROGELBY ENZYMATIC DEGRADATIONa) Calculate the amount of cellulose present in thesample well using the following equation.
NOTE: 100 µl of 1% GrowDex = 1 mg cellulose .
Sample well volume (µl) x % GrowDex
concentration / 100% = cellulose/samplewell (mg)b) Calculate the amount enzyme needed to degrade thecellulose in the sample well using the following equation.
NOTE: 300 µg GrowDase enzyme is required todegrade 1 mg of cellulose.
Amount of cellulose/sample well (mg) x 300µg/mg = Amount of GrowDase enzyme (µg)c) Calculate the volume of GrowDase stock solution
needed using the following equation:
GrowDase amount (µg) / GrowDase stock

concentration (10 µg/µl) = Volume of GrowDase
stock solution (µl)d) Prepare the working concentration of GrowDaseby diluting the stock solution with culture media. The
volume of GrowDase working solution is recommended
to be the same as the volume of GrowDex in the well:
GrowDex volume in well (µl) – Volume (µl) ofGrowDase stock solution (10 mg/ml) = Volumeof culture media for dilution (µl)e) Pipette the diluted GrowDase onto the top of thesample in the microplate.
f) Incubate the plate at 37°C for a minimum of 8 hours
until the hydrogel has fully degraded.
g) Recover the cells from the well using standard
techniques.
Continues

Continued

6. EXAMPLE EXPERIMENTAL PROCEDURESAMPLE: 80 µl of 0.9% GrowDex/cell mix per well ina 96-well microplatec) Volume of GrowDase stock solution needed: 216 µl/10 µg / µl = 21.6 µl GrowDase enzyme stock solutionMATERIALS- GrowDase enzyme, 10 mg/ml (10 µg/µl)- Cell culture mediumd) Prepare the working concentration of GrowDase bydiluting the stock solution (10 mg/ml) with culture media:
80 µl - 21.6 µl = 58.4 µl culture media for dilutione) Pipette the diluted GrowDase onto the top of thesample in the microplate.

METHODa) a) Amount of cellulose present in the sample: 80 µl x0.9%/100% = 0.72 mg cellulose/sample wellf) Incubate the plate at 37°C until the hydrogel has
fully degraded.

b) Amount of GrowDase needed to degrade thecellulose in the sample: 0.72 mg x 300 µg/mg = 216µg GrowDase enzymeg) Recover the cells from the well using standardtechniques.

7. DILUTION TABLE
Volumes of GrowDase enzyme and cell culture medium required for the preparation of 100 µl of GrowDase
working solution (300 µg/mg) for the degradation of 100 µl of GrowDex.

GROWDEXCONCENTRATION IN
100 µL OF SAMPLEAMOUNT OFGROWDASE ENZYMENEEDEDVOLUME OFGROWDASE ENZYME STOCKSOLUTION (10 MG/ML)

VOLUME OF CELLCULTURE MEDIUM

1.0%

300 µg

30 µl

70 µl

0.9%

270 µg

27 µl

73 µl

0.8%

240 µg

24 µl

76 µl

0.7%

210 µg

21 µl

79 µl

0.6%

180 µg

18 µl

82 µl

0.5%

150 µg

15 µl

85 µl

0.4%

120 µg

12 µl

88 µl

0.3%

90 µg

9 µl

91 µl

0.2%

60 µg

6 µl

94 µl

8. ORDERING INFORMATIONCATALOGUE CODEDESCRIPTIONQUANTITY (ml)

900 102 001

GrowDase enzyme

1.25900 102 002

GrowDase enzyme

2.5

100 103 905

GrowDex + GrowDase combo pack

5.0 + 2.5

You can order products online at: www.upmbiomedicals.com/store
Or contact us at biomedicals.sales@upm.com for a quotation or to place an order.

9. CONTACT INFORMATION
Additional information on all products and applications can be found on our website:www.upmbiomedicals.com
Should you have any technical questions regarding this product or its intended use please contact us at:
EMAIL:

biomedicals.support@upm.com

POST:												UPM Biomedicals
Alvar Aallon katu 1
P.O. Box 380
00101 HelsinkiFinlandTEL:					FAX:		

+358 (0)204 15 111
Request to speak with someone from the Biomedicals team+358 (0)204 15 110

Version 2.0 (02/2020)