Abstract
| Original language | English |
|---|---|
| Pages (from-to) | 541-554 |
| Number of pages | 13 |
| Journal | The International Journal of Biochemistry & Cell Biology |
| Volume | 39 |
| Issue number | 3 |
| DOIs | |
| Publication status | Published - 2007 |
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Keratinoycyte serum-free medium maintains long term liver gene expression and function in cultured rat hepatocytes by preventing the loss of liver-enriched transcription factors. / Li, W C; Ralphs, K L; Slack, J; Tosh, D.
In: The International Journal of Biochemistry & Cell Biology, Vol. 39, No. 3, 2007, p. 541-554.Research output: Contribution to journal › Article
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TY - JOUR
T1 - Keratinoycyte serum-free medium maintains long term liver gene expression and function in cultured rat hepatocytes by preventing the loss of liver-enriched transcription factors
AU - Li, W C
AU - Ralphs, K L
AU - Slack, J
AU - Tosh, D
PY - 2007
Y1 - 2007
N2 - Freshly isolated hepatocytes rapidly lose their differentiated properties when placed in culture. Therefore, production of a simple culture system for maintaining the phenotype of hepatocytes in culture would greatly facilitate their study. Our aim was to identify conditions that could maintain the differentiated properties of hepatocytes for up to 28 days of culture. Adult rat hepatocytes were isolated and attached in Williams' medium E containing 10% serum. The medium was changed to either fresh Williams' medium E or keratinocyte serum-free medium supplemented with dexamethasone, epidermal growth factor and pituitary gland extract. The hepatic phenotype was then analysed using RT-PCR, immunohistochemistry, Western blotting and assays of liver function. Cells cultured in keratinocyte serum-free medium supplemented with dexamethasone, epidermal growth factor and pituitary gland extract maintained their phenotype for 3-4 weeks, based on expression of liver proteins, ureagensis and response to xenobiotics. In contrast, hepatocytes cultured in Williams' medium E rapidly lost the expression of liver proteins after 3 days. Cells cultured in keratinocyte serum-free medium supplemented with dexamethasone, epidermal growth factor and pituitary gland extract maintained their expression of liver-enriched transcription factors (C/EBP alpha and beta, HNF4 alpha and RXR alpha) while expression was either lost or reduced in cells cultured in Williams' medium E. These results suggest that keratinocyte serum-free medium supplemented with dexamethasone, epidermal growth factor and pituitary gland extract can maintain the hepatic phenotype for a prolonged period and that this is probably related to the continued expression of the liver-enriched transcription factors. (c) 2006 Elsevier Ltd. All rights reserved.
AB - Freshly isolated hepatocytes rapidly lose their differentiated properties when placed in culture. Therefore, production of a simple culture system for maintaining the phenotype of hepatocytes in culture would greatly facilitate their study. Our aim was to identify conditions that could maintain the differentiated properties of hepatocytes for up to 28 days of culture. Adult rat hepatocytes were isolated and attached in Williams' medium E containing 10% serum. The medium was changed to either fresh Williams' medium E or keratinocyte serum-free medium supplemented with dexamethasone, epidermal growth factor and pituitary gland extract. The hepatic phenotype was then analysed using RT-PCR, immunohistochemistry, Western blotting and assays of liver function. Cells cultured in keratinocyte serum-free medium supplemented with dexamethasone, epidermal growth factor and pituitary gland extract maintained their phenotype for 3-4 weeks, based on expression of liver proteins, ureagensis and response to xenobiotics. In contrast, hepatocytes cultured in Williams' medium E rapidly lost the expression of liver proteins after 3 days. Cells cultured in keratinocyte serum-free medium supplemented with dexamethasone, epidermal growth factor and pituitary gland extract maintained their expression of liver-enriched transcription factors (C/EBP alpha and beta, HNF4 alpha and RXR alpha) while expression was either lost or reduced in cells cultured in Williams' medium E. These results suggest that keratinocyte serum-free medium supplemented with dexamethasone, epidermal growth factor and pituitary gland extract can maintain the hepatic phenotype for a prolonged period and that this is probably related to the continued expression of the liver-enriched transcription factors. (c) 2006 Elsevier Ltd. All rights reserved.
UR - http://dx.doi.org/10.1016/j.biocel.2006.10.017
U2 - 10.1016/j.biocel.2006.10.017
DO - 10.1016/j.biocel.2006.10.017
M3 - Article
VL - 39
SP - 541
EP - 554
JO - The International Journal of Biochemistry & Cell Biology
JF - The International Journal of Biochemistry & Cell Biology
SN - 1357-2725
IS - 3
ER -