摘要
目的 以体外培养表皮干细胞 (ESC)为实验平台 ,观察感觉神经肽P物质 (SP)在ESC分化中的作用。 方法 以黏附分离法分离、纯化新生Wistar大鼠的ESC,在出现ESC克隆生长时加入SP刺激 ,分别于刺激前及刺激后 2 4、4 8、72、96、14 4、192、2 4 0、2 88、336、384、4 32h取样进行角蛋白 14(K14)免疫组织化学染色并用流式细胞仪 (FCM),鉴定细胞分群及比例。 结果 ESC在加入SP后可以继续呈片状聚集生长 ,细胞K14染色阳性 ,提示为短暂扩充细胞 (TAC)。FCM检测结果提示 ,ESC经SP刺激后出现TAC细胞群落 ,细胞比例随时间的延长上升。 结论 SP可以诱导体外培养ESC分化为TAC。
Objective To investigate the effect of sensory neuropeptide substance P(SP) on the differentiation of cultured epidermal stem cells (ESC) in vitro,with in vitro cultured ESC as the platform. Methods ESC from newborn Wistar rats were isolated, purified by repeated passages in culture. SP was added for stimulation when ESC clone grew. Immunohistochemistry staining with K14 antibody, and flow cytometry (FCM) was performed at 0,24th,48th,72nd,96th,144th,192nd,240th,288th,336th,384th,432nd post differentiation hours(PDH) to identify the cell groups and to detect if there were transient amplifying cells(TAC)among the cells. Results ESC in culture formed large colonies after SP treatment with positive staining for K14,indicating that they were TACs. The results of FCM indicated that when ESC were stimulated by SP, TAC colony formation occured and the cell number increased in a constant speed. Conclusion ESC could differentiate into TAC by neuropeptide SP induction, and the numbert of ESC kept on a certain level during the process.
出处
《中华烧伤杂志》
CAS
CSCD
2004年第5期295-298,共4页
Chinese Journal of Burns
基金
国家重点基础研究发展规划专项经费资助项目(G19990 5 42 0 4)
