Repopulation of human origin hepatocyte progenitor-like cell line, THLE-5b, in the SCID mouse liver under p21-mediated cell growth-arresting conditions.
The in vivo repopulation of hepatocytes depends on donor cell growth potential and recipient conditioning. We herein demonstrate the successful cell transplantation of a human hepatocyte cell line, THLE-5b, into the SCID mouse liver by means of a rather mild conditioning using a 55%  hepatectomy and p21 transfection. Adult human liver-derived cells, THLE-5b, are SV40 T antigen-immortalized epithelial cells. A phenotypic examination of THLE-5b showed they expressed hepatic stem cell markers such as EpCAM, OCT3/4, and Thy-1, thus indicating the immature nature of the cells.  A three-dimensional aggregate culture of THLE-5b showed a higher expression level of liver-specific genes such as albumin, α1-antitrypsin, and CYP3A4, thus suggesting that THLE-5b possess the capability to differentiate into hepatocytes. In a cell transplantation experiment, the cell  cycle regulator p21 was transfected with adenoviral vector into the SCID mouse liver. On the next day, 8 × 10(5) cells of GFP-transfected THLE-5b were injected intrasplenically, together with the intraperitoneal administration of anti-asialo GM1 antibodies. The following day,  a partial hepatectomy was performed. The GFP-THLE-5b cells were observed to have migrated and become integrated into the liver parenchyma 14 days after transplantation. The present protocol is thus considered to be a novel experimental model to elucidate the mechanism of hepatocyte repopulation  and to develop efficient stem cell therapy in the liver.
Division for Advanced Medical Services, National Center for Child Health and Development, Setagaya-ku, Tokyo, Japan. senosawa@nch.go.jp
Shin
S
Enosawa
Taisuke
T
Yamazaki
Hitoshi
H
Kohsaka
Takayoshi
T
Tokiwa
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D000951:Antigens, Neoplasm / Q000378:metabolism; D015815:Cell Adhesion Molecules / Q000378:metabolism; D002460:Cell Line; D002465:Cell Movement; D049109:Cell Proliferation; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000235:genetics / Q000378:metabolism; D051544:Cytochrome P-450 CYP3A / Q000378:metabolism; D000071858:Epithelial Cell Adhesion Molecule; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D022781:Hepatocytes / Q000166:cytology* / Q000378:metabolism / Q000637:transplantation; D006801:Humans; D008099:Liver / Q000378:metabolism* / Q000473:pathology; D008297:Male; D051379:Mice; D016513:Mice, SCID; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D012709:Serum Albumin / Q000378:metabolism; D018800:Thy-1 Antigens / Q000378:metabolism; D014162:Transfection; D014183:Transplantation, Heterologous; D000515:alpha 1-Antitrypsin / Q000378:metabolism
D000951:Antigens, Neoplasm; D015815:Cell Adhesion Molecules; D050759:Cyclin-Dependent Kinase Inhibitor p21; C499379:EPCAM protein, human; D000071858:Epithelial Cell Adhesion Molecule; D050814:Octamer Transcription Factor-3; D012709:Serum Albumin; D018800:Thy-1 Antigens; D000515:alpha 1-Antitrypsin; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human