Establishing a serum-free primary culture method for cortical neurons of new-born rats
10.3969/j.issn.1671-8348.2014.29.018
- VernacularTitle:新生大鼠皮层神经元体外无血清原代培养
- Author:
Chuanhao YANG
;
Dong ZHAO
;
Qi LIU
;
Yunxiang JI
;
Lingjian LI
;
Hui XU
;
Jing DAI
;
Xuejun HE
;
Yezhong WANG
- Publication Type:Journal Article
- Keywords:
neuron;
ceus,cultured;
delture media;
cell model
- From:
Chongqing Medicine
2014;(29):3901-3903,3906
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a serum-free primary culture method for cortical neurons of new-born rats .Methods The cortical tissue was digested and the cells were planted in the medium containing 10% fetal bovine serum ,and then maintained feed-ing with neurobasal medium containing B27 after 4 to 8 h .The morphology was observed under phase-contrast microscope .RT-PCR ,Western blot and immunocytochemistry were applied to identify the expression of NSE gene and protein in neurons .Results A large number of neurons began to adhere to the cover glasses after 2 to 8 h .They showed different shapes-shuttle ,triangle pyram-idal ,or no regular after clinging to the plate .Their processes connected to nets and were different in length and thickness .They well developed at the 7th to 10th day .The isolated and cultured cells were confirmed as neurons by RT-PCR ,Western blot and immuno-cytochemistry .Conclusion This technique is an easy and practical tool for primary culture of new-born rats cortical neurons with high purity ,and can be used as an in-vitro model of research .
