Fukumitsu K, Ishii T, Yasuchika K, Amagai Y, Saitoh M, Kawamoto T, Kawase E, Suemori H, Nakatsuji N, Ikai I, Uemoto S
Establishment of a cell line derived from mouse fetal liver that have the character to promote the hepatic maturation of mouse embryonic stem cells by co-culture method. [JOURNAL ARTICLE]
Tissue Eng Part A 2009 Jun 26.
Stromal cells residing in murine fetal livers have the ability to promote the hepatic maturation of murine embryonic stem cells (ESCs) and hepatic progenitor cells (HPCs) in vitro. These stromal cells were isolated as the CD49f(+/-)CD45(-)Thy1(+)gp38(+) cell fraction. The present study established a murine fetal liver stromal cell line that induced hepatic maturation in mouse ESCs and HPCs. A transgene containing a temperature sensitive SV40 large T antigen was transfected into the primary fetal liver stromal cells. These immortalized cells, which were named as the MLSgt cells, induced both mouse ESCs and HPCs to differentiate into mature hepatocyte-like cells using a co-culture method. Since the MLSgt was not a cloned cell line, one clone, MLSgt20, was selected as the line with the character to induced the hepatic differentiation, which was comparable to its parental stromal cells. The ESC-derived endoderm cells co-cultured with the MLSgt-20 cells expressed mature hepatocyte specific gene markers, including glucose-6-phosphatase, tyrosine aminotransferase, tryptophan-2,3-dioxgenase and cytochrome P450 (CYP1a1, Cyp1b1, Cyp1a2, and Cyp3a11). In addition, these cells also exhibited hepatic functions, such as glycogen storage and ammonia metabolism. Transmission electron microscopy showed that the co-cultured ESCs expressed the morphologic features of mature hepatocytes. In conclusion, a cell line was established that have the character to promote the hepatic maturation of mouse ESCs and HPCs by co-culture method.
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