南方医科大学学报2017，Vol.37Issue(3)：317-322,6.DOI:10.3969/j.issn.1673-4254.2017.03.07
大鼠雪旺细胞系RSC96条件培养液对少突胶质前体细胞的体外增殖作用
Conditioned medium from rat RSC96 cells promotes proliferation of oligodendrocyte progenitor cells in vitro
摘要
Abstract
Objective To investigate the effect of conditioned medium from rat RSC96 cells (RSC96-CM) on the proliferation of oligodendrocyte progenitor cells (OPCs) and explore the underlying mechanism.Methods OPCs isolated from the spinal cords of SD rats of embryonic day 15 using immunopanning were treated with RSC96-CM.The proliferation of OPCs was detected using 5-bromo-2'-deoxyuridine (BrdU) incorporation assay.The mRNA expressions of PDGF-AA and bFGF in RSC96 cells were detected using RT-PCR,and their protein concentrations in RSC96-CM were detected with enzyme-linked immunosorbent assay (ELISA).The effects of PDGF-AA and bFGF in RSC96-CM on OPC proliferation and the roles of ERK and JNK signaling pathways in RSC96-CM-induced OPC proliferation were determined by application of their specific inhibitors.Results The percentage of BrdU+ OPCs was significantly increased in response to treatment with RSC96-CM (P＜0.05),reaching the peak level when 50％ RSC96-CM was added in the cell culture.RSC96 cells expressed a substantial amount of PDGF-AA and bFGF mRNAs,and PDGF-AA and bFGF protein concentrations in RSC96-CM were higher than those in a conditioned medium (B104CM) we used previously by 0.87 and 0.92 folds,respectively.Both the specific inhibitor of PDGFR signal pathway (AG1295) and the specific inhibitor of bFGFR signal pathway (PD173074) significantly attenuated RSC96-CM-induced OPC proliferation.The specific inhibitors of ERK signal pathway (U0126) and JNK signal pathway (SP600125) significantly decreased the percentage of BrdU + cells in RSC96-CM-induced OPCs (P＜0.01).Conclusion RSC96-CM can effectively promote OPC proliferation,possibly as a result of PDGF-AA and bFGF secretion by RSC96 cells to activate ERK1/2 and JNK signaling pathways.RSC96-CM can be used as a routine stimulator for promoting OPC proliferation.关键词
雪旺细胞系RSC96条件培养液/少突胶质前体细胞/增殖Key words
RSC96 cells/conditioned medium/oligodendrocyte progenitor cells/cell proliferation引用本文复制引用
齐琦,朱安友,吕合作,胡建国..大鼠雪旺细胞系RSC96条件培养液对少突胶质前体细胞的体外增殖作用[J].南方医科大学学报,2017,37(3):317-322,6.基金项目
国家自然科学基金(81471277) （81471277）
安徽省高等学校自然科学研究重点项目(KJ2016A869,KJ2017A232) （KJ2016A869,KJ2017A232）
安徽省高等学校自然科学研究一般项目(KJ2015B049by)Supported by National Natural Science Foundation of China (81471277). （KJ2015B049by）
