Isolation and Culture of Primary Rat Hepatic Stellate Cells

Xibing Zhang; Fang Liu; Ruichao Wu; Yang Gao; Jianghua Ran

doi:10.47297/wspciWSP2516-252701.20220603

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Isolation and Culture of Primary Rat Hepatic Stellate Cells

Volume 6, Issue 3

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Isolation and Culture of Primary Rat Hepatic Stellate Cells
Isolation and Culture of Primary Rat Hepatic Stellate Cells
The Affiliated Calmette Hospital of Kunming Medical University
Vol. 6, Issue 3, Pages: 8-13(2022)
Published： 2022 ，
DOI：10.47297/wspciWSP2516-252701.20220603
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XIBING ZHANG, FANG LIU, RUICHAO WU, YANG GAO & JIANGHUA RAN. (2022). Isolation and Culture of Primary Rat Hepatic Stellate Cells. Creativity and Innovation, 6(3), 8-13..
DOI：

XIBING ZHANG, FANG LIU, RUICHAO WU, YANG GAO & JIANGHUA RAN. (2022). Isolation and Culture of Primary Rat Hepatic Stellate Cells. Creativity and Innovation, 6(3), 8-13.. DOI： 10.47297/wspciWSP2516-252701.20220603.

摘要

Abstract

Improve the rat hepatic stellate cells (HSCs) isolation method to offer a stable and effective cell model for fundamental liver research.Adult SD rats were used

and liver cell suspensions were produced by in situ and ex vivo perfusion

followed by gradient density centrifugation to yield rat HSCs. Trypan blue staining was used to test cell viability

autofluorescence under a fluorescence microscope was used to measure HSC purity

and HSCs were identified using immunofluorescence cytochemical labeling.The output of rat HSCs was greater than 2.0×107

the purity of HSCs was greater than 90%

and the HSC survival rate was greater than 80%. The antigens Desmin and α-smooth muscle actin (α-SMA) were found in immunofluorescence cytochemical labeling of HSCs. Through repeated investigation and comparison

a set of procedures for isolating and growing rat HSCs with easy operation and consistent outcomes was developed in this work.

关键词

Keywords

HSCsCell isolationα-SMADesmin

references

A Geerts. "History, heterogeneity, developmental biology, and functions of quiescent hepatic stellate cells"[J].Semin Liver Dis.2001,21(3):311-35.[2] Takuma Tsuchida, Scott L Friedman. "Mechanisms of hepatic stellate cell activation"[J]. Nat Rev Gastroenterol Hepatol.2017,14(7):397-411. [3] Haruki Senoo, Yoshihiro Mezaki, Mutsunori Fujiwara. "The stellate cell system (vitamin A-storing cell system)"[J]. Anat Sci Int.2017,92(4):387-455.[4] Li Y, Cai S, Zhang L, Li X. "Simultaneous isolation and primary culture of rat hepatocytes, hepatic stellate cells, Kupffer's cells and hepatic sinus endothelial cells"[J]. Nan Fang Yi Ke Da Xue Xue Bao. 2014,34(4):532-7.[5] Ralf Weiskirchen, Axel M Gressner. "Isolation and culture of hepatic stellate cells"[J].Methods Mol Med.2005,117:99-113.

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