Cell Culture

Hepatocyte Culture

Sigma is proud to offer a selection of media, many of which are serum-free, for the in vitro culture of human primary cells, such as keratinocytes, endothelial cells, and epithelial hepatocytes. These primary cell types have potential commercial value in a wide range of emerging technologies and therapeutic applications. Among these are: tissue engineering applications, such as wound healing and burn treatment; high throughput screening for drug toxicology and efficacy studies; gene therapy; pseudo-organs (e.g.. artificial liver) and as model cell systems for developing new therapeutic approaches to human diseases.




Hepatocytes

   Hepatocyte Cell Culture System
Hepatocyte Cell Culture Medium
Hepatocyte Cell Culture Companion Products and Supplements

Hepatocyte Cell Culture System

Hepatocytes are epithelial cells found in the liver. They perform important functions, such as helping to detoxify blood, and to synthesize transport proteins, such as lipoprotein, albumin and transferrin.  Primary and secondary cultures of hepatocytes are useful for studying the mechanisms of liver regeneration and differentiation.  Historically, primary hepatocytes have exhibited a limited replicating lifespan in culture.  In addition, when stimulated to divide in culture they have generally lost differentiated functions such as the ability to synthesize and secrete albumin and transferrin.  While substantial progress has made in understanding the factors which affect these characteristics of hepatocytes in culture, primary and secondary hepatocytes are still not suitable for use in some emerging medical technologies and applications.  Among the emerging potential applications for cultured hepatocytes are gene therapy, bio-artificial organs, cell transplants, drug production, and drug and chemical testing.  These technologies may benefit from the use of cloned and immortalized hepatocytes.  Hence, there is a significant effort to develop hepatocyte cell lines for this purpose.

Hepatocyte Cell Culture Medium

Many classical media (see web-pages in our "Classical Media" section) have been used to culture epithelial hepatocytes. Some examples of these are; Liebovitz L-15, DMEM/F-12, RPMI 1640, Waymouth's MB 752/1 and Williams Medium E.  Growth of hepatocytes in these media requires the use of of serum.

Our most advanced formulation for culturing primary, secondary, and cloned immortalized epithelial hepatocytes is H 1777, which is an enriched variation of Leibovitz L-15 formulation.  It may be successfully used with reduced concentrations of FBS.


Product # Product Name Description Application
H 1777 Hepatocyte Medium
  • Liquid,
  • Proprietary Mixture;
  • With HEPES and sodium bicarbonate buffers;
  • Without L-glutamine;
  • Sterile filtered;
  • Endotoxin tested;
  • Cell culture tested.

H1777 is an enhanced medium formulation based on the original Liebovitz, L-15 medium. It is formulated to support the  proliferation of primary, secondary and immortalized hepatocytes.

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Hepatocyte Cell Culture Companion Products and Supplements

The culture of epithelial hepatocyte cells is supported by a series of products offered by Sigma.  The table below provides a partial list of Sigma products recommended for this purpose.  Also, see the "Hepatocyte Cell Culture Media" table above.


Product # Product Name Description Application
C 6295 Cell Freezing Medium-DMSO, serum-free
  • Liquid
  • Contains minimum essential medium (MEM), dimethyl sulfoxide (8.7%) and methycellulose
  • Sterile filtered
  • Endotoxin tested
  • Cell culture tested

Complete ready to use medium for the cryopreservation of cells.

C 2799 Collagenase
  • Powder
  • High Purity, Type VII
  • Activity: 1000-3000 collagen digesting units/mg solid; 4-12 FALGPA hydrolysis units/mg solid.
  • Contaminating activity: < 1 unit neutral protease and < 1 unit clostripain/mg solid
  • Cell culture tested

Useful for the isolation of epithelial hepatocytes when used in liver perfusion solutions.

D 1435 Dimethyl Sulfoxide, USP
  • Liquid
  • USP tested

DMSO has been shown to stimulate the redifferentiation of dedifferentiated epithelial hepatocytes in vitro.

H 0887 HEPES Buffer, (1M)
  • Liquid
  • Prepared from HEPES free acid and tissue culture grade water
  • Sterile filtered
  • Endotoxin tested
  • Cell culture tested

Useful for the isolation of epithelial hepatocytes when used in liver perfusion solutions.

H 3784 HEPES, sodium salt
  • Powder
  • Purity: > 99.5%, DNase, RNase and protease non detected, < 5 ppm heavy metals and < 5 ppm iron
  • pKa = 7.31 at 37°C; useful pH range 6.8-8.2
  • Endotoxin and bioburden tested
  • Cell culture tested

Useful for the isolation of epithelial hepatocytes when used in liver perfusion solutions.

H 3884 Hyaluronidase
  • Powder, lyophilized
  • Source: Bovine testes
  • High Purity, Type IV-S
  • M. Wt. = 56 kDa
  • Randomly cleaves glycosidic bonds in hyaluronic acid, chondroitin, and chondroitin sulfates
  • Activity: 750-1,500 units/mg solid.  Assayed per the USP XXIV-NF XVII combined edition.
  • Contaminating activity: < 1 unit neutral protease and < 1 unit clostripain/mg solid
  • Cell culture tested

Useful for the isolation of epithelial hepatocytes when used in liver perfusion solutions.

     

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