Purification and culture of astrocytes from cerebral cortex of neonatal rats in vitro
Ma, Y.; Li, W.; Li, Z.; Lü, X.; Wang, X.; Huang, G.
Wei Sheng Yan Jiu 48(5): 795-798
2019
ISSN/ISBN: 1000-8020 PMID: 31601320 Document Number: 697817
To establish an effective method of primary and passage cultured cerebral cortical astrocyte of SD neonatal rat in vitro. Cerebral cortex of two 2-day-old SD rats were taken with aseptic operation and then were cut to pieces. After stripped the pia mater, digested by 500 μL 0. 25% trypsin at 37 ℃ for 15 min. Next, dispersed cell suspension was made by mechanical method and filtered. Cell suspensions were incubated in an uncoated culture bottle at 37 ℃ for 15 min. The cells were inoculated at 5×10~6/m L in the T75 culture flask coated with L-polylysine. The cells were shaken at 200 r/min 37 ℃ for 18 h, then added 1 m L of trypsin to digest cells and then collected the cells. The morphology of the passage cells was observed under inverted phase contrast microscope, and the purity of astrocytes was identified by immunofluorescence staining of GFAP. The proliferate activity of passage cells was determined by MTS assay. The purity of astrocytes was( 97. 86 ± 0. 91) %, and the growth and proliferation activity of astrocytes were good after passage. A rapid, economical and effective method for obtaining astrocytes in the cerebral cortex of newborn rats was established.