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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
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 |
|
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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