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研究発表を行った学会；
・第３４回　日本分子生物学会年会
　２０１１年１２月１３日〜１６日（横浜）

タイトル；Optimization of long-term culture condition for murine primary hepatocytes.

発表者；田山　慎二　氏
　　　（熊本大学　発生医学研究所　多能性幹細胞分野）
Abstract；
Primary culture of hepatocytes has been used as research material for studies of the basic biology of hepatic function and diseases, as well as for toxicity assays in drug development. Human embryonic stem (ES) cells and induced pluripotent stem (iPS) cells are expected as an alternative source for hepatocytes.
To date, many studies have been focused on the establishment of an efficient differentiation procedure to derive hepatic cells from human ES/iPS cells. Like primary hepatocytes, differentiated cells from ES cells might be short-lived and inclined to significantly reduce their activities in culture. In this study we aim to establish a culture system to enable a long-term culture of mouse primary hepatocytes, which would be applicable to ES/iPS cells-derived mature hepatic cells.
Hepatocytes were prepared from 6-week old ICR mice with two step perfusion. We cultured isolated primary hepatocytes and compared with different media or scaffolds, and scored their function by measuring albumin secretion by ELISA, and microscopic observations of the morphological features of hepatocytes. We confirmed that mouse primary hepatocytes could be cultured for a certain period with chemically defined media, without fetal bovine serum (FBS). Scaffold conditions were also optimized by comparing different substrata using the above mentioned FBS free media.
We then tested the optimized culture conditions on hepatocytes differentiated from human iPS cells. These cells turned out to be able to survive for 15 days with the above optimized condition. Taken together, our study demonstrates that it is feasible to culture differentiated hepatocytes under chemically defined conditions.