Spinal cord-derived neural stem cells cultured by serum-free suspension method: separation, cultivation and identification

doi:10.3969/j.issn.2095-4344.2015.50.022

Abstract

Abstract:

BACKGROUND: Until now, there is yet no complete recovery from spinal cord injury in terms of structure and functional recoveries. Neurotrophic factors have limited effects on nerve regeneration. Currently, stem cell transplantation may be an effective way to repair spinal cord injury.
OBJECTIVE: To separate, cultivate and purify mouse spinal cord-derived neural stem cells using serum-free suspension method followed by morphological observation, immunofluorescence technology and multi-lineage differentiation experiments.
METHODS: By using the suspension culture method, mouse spinal cord-derived neural stem cells at embryonic day 13.5 were cultured and purified. Cell morphology changes were observed under inverted microscope. Cell proliferation ability was detected using cell counting kit-8. Nestin and Sox2 expression was detected by immunofluorescence technology. Multilineage differentiation of spinal cord-derived neural stem cells at passage 4 was detected by natural differentiation method in order to prove the differentiation ability.
RESULTS AND CONCLUSION: Serum-free medium suspension culture method was successfully applied to separate spinal cord-derived neural stem cells. Cultured cells had good proliferative ability and highly expressed Nestin and Sox2 that was in accordance with the results of DAPI nucleus staining, suggesting the high purity of cells. After induction, the cells could express both Tuj1 and GFAP, indicating the cells had good differentiation
potential. This experiment has successfully established the isolation, culture, identification system of spinal cord-derived neural stem cells, providing experimental basis for subsequent studies of neural stem cells.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

CLC Number:

R394.2

Li Jun, Xu Da-bo, Fu Qiang, Liu Yan-bin. Spinal cord-derived neural stem cells cultured by serum-free suspension method: separation, cultivation and identification[J]. Chinese Journal of Tissue Engineering Research, 2015, 19(50): 8161-8166.

Figures/Tables 6

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