H2.35

Provides modal system for studying hepatocyte differentiation because some hepatocyte phenotypes are inducible in the cells. Transfection recipient for assays of genetic regulatory sequence function for liver genes that do not function in other cells.

From a primary hepatocyte culture of 6 week old female BALB/c mouse transformed with tsA255 (temperature-sensitive mutant of SV40 virus). H2.35 is temperature sensitive for expression of serum albumin mRNA. During routine propagation at 33°C it exhibits extremely low levels of albumin transcription and mRNA, while at the restructure temp of 39°C albumin mRNA increases 100 fold if the cells are grown in serum free medium on a collagen substratum. Undergoes morphological changes in response to extracellular matrix gel substrata, albumin gene transcription is induced. Liver-enriched transcription factors are induced in serum-free medium and on collagen gels. The cells should be handled under laboratory containment level 2.

Mouse Hepatocyte, SV40 (tsA255) transformed

DMEM (containing 1g / l glucose) + 2mM Glutamine + 250nM Dexamethasone (DXMT) + 4% Foetal Bovine Serum (FBS).

Split sub-confluent cultures (70-80%) 1:6 to 1:10 i.e. seeding at 2-4x10,000 cells/cm² using 0.25% trypsin or trypsin/EDTA; 5% CO₂; 33°C.

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