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[Primary culture and purification of cerebral astrocyte of tree shrew.] [Article in Chinese]

GONG Min, LI Shu-Qing^*, LI Fan

Department of Pathophysiology, Kunming Medical College, Kunming 650031, China

Abstract

The aim of this study is to establish a method to culture and purify cerebral astrocyte of tree shrew (Tupaia belangeri), a kind of new laboratorial animal which is a relative of primates. Newborn tree shrews were used in this experiment. The cortex of cerebrum was isolated and placed in 4°C for 20 min to injure neurons. The cortical tissue was disaggregated by trypsin digestion. Differential attachment method was used to remove fibroblasts. The mixed culture was rinsed by trypsin (0.005%) solution to remove neurons. Upon reaching 70% confluence, the culture was subjected to static trypsin digestion until a white slice film exfoliated from the bottom of culture bottle. This film, i.e. astrocyte layer, was taken out and cultured, and the third passage was identified by immunocytochemical staining and immunofluorescence with anti-glial fibrillary acidic protein (GFAP) antibody. The result showed the purity of tree shrew astrocytes was more than 98%. Thus the method to culture highly purified astrocyte of tree shrew was successfully established, which would contribute to further study in central nervous system physiology and diseases in this new laboratorial animal.

Key words: tree shrew; astrocyte; purification; immunohistochemistry; immunofluorescence

Received: 2010-06-25　　Accepted: 2010-12-15

Corresponding author: 李树清　　E-mail: shuqing591@hotmail.com

Citing This Article：

GONG Min, LI Shu-Qing, LI Fan. [Primary culture and purification of cerebral astrocyte of tree shrew.] [Article in Chinese]. Acta Physiol Sin 2011; 63 (1): 89-92 (in Chinese with English abstract).