RRC ID 11967
Author Fukumitsu K, Ishii T, Yasuchika K, Amagai Y, Kawamura-Saito M, Kawamoto T, Kawase E, Suemori H, Nakatsuji N, Ikai I, Uemoto S.
Title Establishment of a cell line derived from a mouse fetal liver that has the characteristic to promote the hepatic maturation of mouse embryonic stem cells by a coculture method.
Journal Tissue Eng Part A
Abstract 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) 3848 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 gp38-positive and Thy1-positive murine liver stromal (MLSgt) cells, induced both mouse ESCs and HPCs to differentiate into mature hepatocyte-like cells using a coculture method. Since MLSgt is not a cloned cell line, one clone, MLSgt20, was selected as a line with the characteristic to induce hepatic differentiation, which was comparable to its parental stromal cells. The ESC-derived endoderm cells cocultured with the MLSgt20 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 cocultured ESCs expressed the morphologic features of mature hepatocytes. In conclusion, a cell line was established that has the characteristic to promote the hepatic maturation of mouse ESCs and HPCs by a coculture method.
Volume 15(12)
Pages 3847-56
Published 2009-12
DOI 10.1089/ten.TEA.2009.0357
PMID 19558317
MeSH Animals Antigens, Polyomavirus Transforming / metabolism Cell Differentiation* Cell Line / cytology* Coculture Techniques / methods* Embryonic Stem Cells / cytology* Embryonic Stem Cells / metabolism Embryonic Stem Cells / ultrastructure Fetus / cytology* Gene Expression Regulation Liver / cytology* Liver / embryology* Liver / metabolism Membrane Glycoproteins / metabolism Mice Mice, Inbred C57BL Reverse Transcriptase Polymerase Chain Reaction Stromal Cells / cytology Stromal Cells / metabolism Thy-1 Antigens / metabolism Transfection alpha-Fetoproteins / metabolism
IF 3.508
Times Cited 6
WOS Category BIOTECHNOLOGY & APPLIED MICROBIOLOGY CELL & TISSUE ENGINEERING CELL BIOLOGY
Resource
DNA material pSVtsA58 ori (-) SV40 ori (-) DNA cDNA (RDB01126)