Transient Expression of Pax4 in Murine Embryonic Stem Cell Promotes Differentiation of Insulin-producing Cell and Hepatocyte-like Cell

• Main Authors: Ya-Hui Chang, 張雅惠
• Other Authors: Hung-Hai Ku
• Format: Others
• Language: en_US
• Published: 2005
• Online Access: http://ndltd.ncl.edu.tw/handle/17246300702731144017

碩士 === 國立陽明大學 === 解剖暨細胞生物學研究所 === 93 === Embryonic stem (ES) cells with unlimited dividing capacity can be induced to differentiate into cells of all three primary germ layers including endodermal insulin-producing β cells and hepatocyte. They are expected to be useful for tissue engineering and cell replacement therapy (CRT). In this study, the constitutive enhanced green fluorescent protein (EGFP)-expressing murine embryonic stem (mES) cell line ESC26GJ was engineered to express pax4 transiently (named Pax4+ mES cells) leads to insulin-producing cell clusters production. In the pax4-transfected insulin-producing cells, Insulin 1, Insulin 2 and pdx1 genes expressed significantly as compared with the differentiated cells derived from the parental ESC26GJ. Furthermore, Ngn3 and several genes necessary for glucose-induced insulin secretion, Kir6.2, glucokinase (GK) and the glucose transporter 2 (GLUT2) gene expressions were also detected. There are about 42% insulin-producing cells with the immunohistochemical analysis. In addition, certain endoderm marker genes expressed in the early stage during in vitro pancreatic differentiation means that these cells are endoderm-like cells. To investigate the potential of pax4-transfected endoderm-like cells to transdifferentiate into other endodermal cell-lineages, a modified hepatic induction protocol from Hamazaki et al. in 2001 was used. As we expected, liver-specific genes expression was detected in the differentiated cells at the terminal differentiation stage, and the cell morphology of hepatocyte-like cells was similar to hepatocytes, too. Altogether, this study provides an induction protocol to obtain endodermal insulin-producing cell clusters and hepatocyte-like cells from mES cells.