A new immortalized rat cell line, hepatic stellate cell-PQ, exhibiting characteristics of hepatic stellate cell.
BACKGROUND
Playing a central role in hepatic fibrosis, hepatic stellate cell (HSC) has made itself the major target of research. The limited supply of HSC, however, can not meet the ever growing need of experiment. Establishment and identification of novel immortalized HSC line thus may be urgently required.


METHODS
Primary HSCs were isolated from a normal adult male Sprague-Dawley rat by in situ perfusion with collagenase IV and pronase E, and then were purified by single-step density gradient centrifugation with nycodenz. Once they reached total activation in culture, a new immortalized myofibroblast-like HSC line was established through cellular cloning. Its characteristics were identified by means of immunocytochemical staining, light microscopy, transmission electron microscopy, and growth curve analysis.


RESULTS
The novel HSC line, termed HSC-PQ, had a doubling time of about 75 hours in the Dulbecco's modified Eagle medium (DMEM) containing 20% fetal bovine serum. Most of the main morphological characteristics of the differentiated primary HSC could be detected in HSC-PQ cell, while functional features of activated HSC such as alpha-smooth muscle actin, desmin, collagen type I, collagen type III, fibronectin, laminin and other extracellular matrix proteins could also be found in it except for collagen type IV. In contrast, fat droplets and autofluorescence of vitamin A disappeared in the HSC-PQ line. This cell line had been maintained in culture for over 30 passages and more than 1 year with little alternation in biological characteristics.


CONCLUSIONS
A new rat HSC line (HSC-PQ) has been successfully established. It consistently retains the characteristics of activated primary HSC, and has proved to be immortalized.
Pan
Qin
Q
Department of Gastroenterology, Xinhua Hospital, Shanghai Second Medical University, Shanghai 200092, China. fangchunhua@online.sh.cn
Li
Ding-Guo
DG
Lu
Han-Ming
HM
Wang
Yu-Qing
YQ
Zhang
Wen-Zhu
WZ
Xu
Qin-Fang
QF
D000704:Analysis of Variance; D000818:Animals; D017209:Apoptosis / Q000502:physiology; D002460:Cell Line; D002461:Cell Line, Transformed; D049109:Cell Proliferation; D005434:Flow Cytometry; D022781:Hepatocytes / Q000166:cytology* / Q000502:physiology*; D007150:Immunohistochemistry; D008103:Liver Cirrhosis / Q000473:pathology / Q000503:physiopathology; D008297:Male; D008855:Microscopy, Electron, Scanning; D011336:Probability; D051381:Rats; D017207:Rats, Sprague-Dawley; D012680:Sensitivity and Specificity; D020858:Tissue and Organ Harvesting