Establishment of an immortalized porcine liver cell line JSNK-1 with retroviral transduction of SV40T.
Maintenance of freshly isolated porcine liver cells in vitro is limited for a short period of time. Therefore, establishment of easy handling cell lines is extremely important for in vitro study for liver cells and their possible utilization for cell differentiation and growth of stem cells. Porcine liver cells were transduced with a retroviral vector SSR#69 expressing SV40T, one of SSR#69-immortalized porcine liver cell lines, JSNK-1, was established and characterized. Morphology of JSNK-1 cells was spindle shaped. When the cells became confluent, JSNK-1 cells revealed hills-and-valleys pattern. In the presence of vitamin A, JSNK-1 cells showed big droplets inside the cytoplasm, which were positive with PAS staining. JSNK-1 cells showed the gene expression of collagen type 1α1, collagen type 1α2, FLT-1, β-actin, and SV40T. Immunostaining study revealed that JSNK-1 cells produced collagen, vimentin, and α-smooth muscle actin. JSNK-1 cells possessed the characteristics of the liver stellate cells. JSNK-1 cells produced hepatocyte growth factor (HGF) in a time-dependent manner. When cocultured with iPS cells towards the hepatic differentiation, JSNK-1 cells facilitated their hepatic differentiation in terms of albumin production. In conclusion, JSNK-1 cells would be valuable in the study of liver stellate cell pathophysiology and contribute to the optimization of hepatic differentiation of iPS cells.
Shahid
Javed M
JM
Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine and Dentistry, Okayama, Japan. drshjaved@gmail.com
Iwamuro
Masaya
M
Sasamoto
Hiromi
H
Kubota
Yasuhiro
Y
Seita
Masayuki
M
Kawamoto
Hironobu
H
Nakaji
Shuhei
S
Noguchi
Hirofumi
H
Yamamoto
Kazuhide
K
Kobayashi
Naoya
N
D000418:Albumins / Q000096:biosynthesis; D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000378:metabolism*; D001681:Biological Assay; D015415:Biomarkers / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D002461:Cell Line, Transformed / Q000166:cytology* / Q000648:ultrastructure; D049109:Cell Proliferation; D048430:Cell Shape; D018920:Coculture Techniques; D003094:Collagen / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D005786:Gene Expression Regulation; D055166:Hepatic Stellate Cells / Q000166:cytology / Q000378:metabolism; D017228:Hepatocyte Growth Factor / Q000096:biosynthesis; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism; D008099:Liver / Q000166:cytology*; D051379:Mice; D012190:Retroviridae / Q000235:genetics*; D034421:Sus scrofa; D014161:Transduction, Genetic; D014746:Vimentin / Q000378:metabolism
D000418:Albumins; D000952:Antigens, Polyomavirus Transforming; D015415:Biomarkers; D014746:Vimentin; D017228:Hepatocyte Growth Factor; D003094:Collagen