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CM3000 — Thawing/Plating Supplement Pack

For use with:  Williams E Medium (WEM, A1217601)

Applications: Thawing and plating cryopreserved hepatocytes, plating fresh hepatocytes

Contents: (concentrations in parenthesis) Final concentrations:
Fetal Bovine Serum (5%)
Dexamethasone in DMSO (10 mM) (1 μM)
Thawing/Plating Cocktail — A, containing (18 mL total volume):  
  • 5.0 mL Penicillin/Streptomycin (10,000 U/mL / (10,000 μg/mL)
(1%)
  • 500 μL Human Recombinant Insulin (4 mg/mL)
(4 μg/ml)
  • 5.0 mL GlutaMAX™ (200 mM/100 X)
(2 mM)
  • 7.5 mL HEPES, pH 7.4 (1M)
(15 mM)

Directions for supplementing medium * :

To 500 ml medium, add:
  • Entire contents of Fetal Bovine Serum tube (25 mL)
  • 50.0 μL Dexamethasone in DMSO
  • Entire contents of Thawing/Plating Cocktail — A (18 mL)

CM4000 — Cell Maintenance Supplement Pack

For use with: Williams E Medium (WEM, A1217601)

Applications:
Incubation medium for cells in suspension, maintenance of cultured hepatocytes

Contents: (concentrations in parenthesis) Final concentrations:
Dexamethasone in DMSO (10 mM) (0.1μM)
Cell Maintenance Cocktail — B, containing (20 mL total volume):  
  • 2.5 mL Penicillin/Streptomycin (10,000 U/mL / (10,000 μg/mL)
(0.5%)
  • 5.0 mL ITS+

  • human recombinant insulin (0.625 mg/mL)
  • human transferrin (0.625 mg/mL)
  • selenous acid (0.625 μg/mL)
  • bovine serum albumin (BSA) - (0.125 g/mL) 
  • linoleic acid (0.535 mg/mL)



(6.25 μg/ml)
(6.25 μg/ml)
(6.25 ng/mL)
(1.25 mg/ml)
(5.35 μg/mL)
  • 5.0 mL GlutaMAX™ (200 mM/100 X)
(2 mM)
  • 7.5 mL HEPES, pH 7.4 (1M)
(15 mM)


Directions for supplementing medium * :

To 500 ml medium, add:

  • 5.0 μl Dexamethasone in DMSO
  • Entire contents of Cell Maintenance Cocktail — B (20 mL)

* To extend life of these products, we recommend supplementing small aliquots of medium just prior to use. Remaining medium, cocktail or dexamethasone should be stored at 4°C for up to twelve months or up to expiration date listed on bottle. Remaining serum should be stored at -20°C.

Table 1 — Volumes of cocktail A and supplements for supplementing aliquots.

WEM Cocktail A FBS Dex
50 mL 1.8 mL 2.5 mL 5 μL
100 mL 3.6 mL 5 mL 10 μL
200 mL 7.2 mL 10 mL 20 μL
300 mL 10.8 mL 15 mL 30 μL
400 mL 14.4 mL 20 mL 40 μL
500 mL 18 mL 25 mL 50 μL



Table 2:  Volumes of cocktail B and supplements for supplementing aliquots.

 WEM Cocktail B Dex
 50 mL 2 mL 0.5 μL
100 mL 4 mL 1 μL
200 mL 8 mL 2 μL
300 mL 12 mL 3 μL
400 mL 16 mL 4 μL
500 mL 20 mL 5 μL

 

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 LT131                     rev 1-Mar-2011