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2005•Chinese Journal of Gastroenterology and HepatologyRequires access

The improved method of isolating and identifying rat hepatic stellate cells

Yuanyi Zheng

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Abstract

Objective To investigate more economical and steadier method for the isolating and culturing of hepatic stellate cells (HSCs). Methods The method for isolating of normal rat HSC was established with collagenase in situ liver recirculation perfusion, and 18% Nyco denz density gradient centrifugation. HSCs were trained in digestion DMEM culture medium. De smin, dt-SMA and ICAM-1 were examined for identifing the cells. Results The quantity of HSC was above 3.5～10~7 cells per rat. The purity and the viability of the cells were about 96～98 percent. Conclusion The result showed that this method of isolating HSC was more economical and steadier; the relatively stead y cell line was acquired by subculturing.

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What this paper is about

Objective To investigate more economical and steadier method for the isolating and culturing of hepatic stellate cells (HSCs). Methods The method for isolating of normal rat HSC was established with collagenase in situ liver recirculation perfusion, and 18% Nyco denz density gradient centrifugation. HSCs were trained in digestion DMEM culture medium. De smin, dt-SMA and ICAM-1 were examined for identifing the cells. Results The quantity of HSC was above 3.5～10~7 cells per rat. The purity and the viability of the cells were about 96～98 percent. Conclusion The result showed that this method of isolating HSC was more economical and steadier; the relatively stead y cell line was acquired by subculturing.

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Available abstract

Objective To investigate more economical and steadier method for the isolating and culturing of hepatic stellate cells (HSCs). Methods The method for isolating of normal rat HSC was established with collagenase in situ liver recirculation perfusion, and 18% Nyco denz density gradient centrifugation. HSCs were trained in digestion DMEM culture medium. De smin, dt-SMA and ICAM-1 were examined for identifing the cells. Results The quantity of HSC was above 3.5～10~7 cells per rat. The purity and the viability of the cells were about 96～98 percent. Conclusion The result showed that this method of isolating HSC was more economical and steadier; the relatively stead y cell line was acquired by subculturing.

Key concepts: Hepatic stellate cell, Collagenase, Differential centrifugation, Centrifugation, Cell culture, Digestion (alchemy), Perfusion, Molecular biology

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