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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；ALB gene targeting in human iPS and ES cells with helper-dependent adenoviral vector for monitoring hepatic differentiation.

発表者；梅田　香穂子　氏
　　　（熊本大学　発生医学研究所　多能性幹細胞分野）
Abstract；
The use of human ES or iPS cell derived hepatocyte will enable evaluation of toxicity at an earlier stage of drug development. We have established a hepatic differentiation procedure in vitro (Shiraki et al., 2008). However, differentiation efficiency is still low. Furthermore, we have to kill the differentiated cells for RT-PCR and immunostaining for estimation of differentiation efficiency.
We aim at the establishment and characterization of the Albumin (ALB)-mKO1 (monomeric kusabira orange) knock-in human ES and iPS cell lines, which enable monitoring of the differentiation of the hepatocyte from human ES and iPS cells in living cells. Gene targeting was carried out using HDAdV (Helper dependent adenovirus vector) system, which was shown as an efficient approach to genetically manipulate human ES cells. The ALB-mKO1 knock in cells were used to visualize hepatic differentiation in vitro. The expression of mKO1 reporter recapitulated the endogenous ALB gene expression. Isolated mKO1-expressing cells by flow cytometry were confirmed to be of hepatic characteristics in that they expressed transcripts of the molecular markers of the mature hepatocytes.
In conclusion, these ALB-mKO1 knock-in human ES and iPS cell lines will be a powerful tool to monitor specific cell lineage in living cells, screen the drugs which potentiates an efficient induction, for quantifying and purifying specific cells by flow cytometry.