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.

WEMCocktail AFBSDex
50 mL1.8 mL2.5 mL5 μL
100 mL3.6 mL5 mL10 μL
200 mL7.2 mL10 mL20 μL
300 mL10.8 mL15 mL30 μL
400 mL14.4 mL20 mL40 μL
500 mL18 mL25 mL50 μL



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

 WEM Cocktail B Dex
 50 mL2 mL0.5 μL
100 mL4 mL1 μL
200 mL8 mL2 μL
300 mL12 mL3 μL
400 mL16 mL4 μL
500 mL20 mL5 μL

 

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 LT131                     rev 1-Mar-2011