Isolation and culture of hepatic stellate cells from mouse liver

• Published in: Acta biochimica et biophysica Sinica Vol. 46; no. 4; pp. 291 - 298
• Main Authors: Chang, Wenju, Yang, Mengxuan, Song, Lujun, Shen, Kuntang, Wang, Hongshan, Gao, Xiaodong, Li, Min, Niu, Weixin, Qin, Xinyu
• Format: Journal Article
• Language: English
• Published: China 01.04.2014
• Subjects: Animals; Cells, Cultured; Hepatic Stellate Cells - cytology; Kupffer Cells - cytology; Mice; Mice, Inbred C57BL; 分离培养; 小鼠; 文化; 磷酸盐缓冲液; 细胞存活率; 肝星状细胞; 肝脏; 造血干细胞; cell purification; cell culture; dichloromethylene diphosphate; hepatic stellate isolation; liposome
• ISSN: 1672-9145; 1745-7270
• DOI: 10.1093/abbs/gmt143

Hepatic stellate cells （HSCs） are the primary extracellular matrix-producing cells within the river and have numerous vital functions. A robust protocol for the isolation and culture of HSCs is important for further investigations of cell functions and related mechanisms in river disease. The volume of the mouse river is much smaller than that of the rat river, which makes it much more difficult to isolate mouse HSCs （mHSCs） than rat HSCs. At present, isolating mHSCs is still a challenge because there is no efficient, robust method to isolate and culture these cells. In the present study, C57BL/6J mice were intravenously injected with riposomeencapsulated dichloromethylene diphosphate （CL2MDP） to selectively eliminate Kupffer cells from the river. The mouse livers were then perfused in situ, and the mHSCs were isolated with an optimized density gradient centrifugation technique. In the phosphate buffer solution （PBS）-liposome group, the yield of mHSCs was （1.37 ±0.23） × 10^6/g river, the cell purity was （90.18 ± 1.61）%, and the cell survival rate was （94.51 ±1.61）%. While in the CL2MDP-liposome group, the yield of mHSCs was （1.62 ±0.34）× 10^6/g liver, the cell purity was （94.44 ± 1.89）%, and the cell survival rate was （94.41 ±1.50）%. Based on the yield and purity of mHSCs, the CL2MDP-riposome treatment was superior to the PBS-liposome treatment （P 〈 0.05, P 〈 0.01）. This study established successfully a robust and efficient protocol for the separation and purification of mHSCs, and both a high purity and an adequate yield of mHSCs were obtained.