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作 者:王晓玲[1] 汪涛[1] 汪雅妮[1] 王学谦[2]
机构地区:[1]天津中医药大学中医学院组织学与胚胎学教研室,天津300193 [2]天津医院,天津300193
出 处:《中国中西医结合杂志》2012年第1期93-96,共4页Chinese Journal of Integrated Traditional and Western Medicine
基 金:天津市高等学校科技发展基金计划项目(No.20060301);国家自然科学基金面上项目(No.30772881)
摘 要:目的观察在体外造血微环境中,当归多糖(angelica polysaccharides,APS)对不同培养条件下乳鼠骨骼肌卫星细胞(muscle satellite cells,MSCs)增殖及干细胞因子受体蛋白(c-kit)表达的影响。方法原代培养MSC,5天后采用结蛋白(desmin)免疫细胞化学鉴定;细胞随机分为8组,即空白对照组,骨髓基质细胞培养上清组,加入含100、200、300μg/mLAPS的DMEM/F12培养基实验组及经100、200、300μg/mLAPS干预后骨髓基质细胞条件培养基组,采用MTT法检测各组细胞的增殖活性及对细胞进行c-kit免疫细胞化学染色。结果分离培养的MSCs呈desmin染色阳性;MTT法检测发现各条件培养基实验组MSCs增殖显著;c-kit免疫强阳性反应存在于条件培养基培养的各组MSCs中,c-kit主要表达于细胞质。结论经APS干预的骨髓基质细胞条件培养基可以有效改变MSCs的生长特性,明显促进MSCs增殖及c-kit的表达,c-kit在MSCs增殖过程中可能起某种调节作用。Objective To observe the effects of angelica polysaccharides (APS) on the proliferation of mouse skeletal muscle satellite cells (MSCs) and c-kit expression in different in vitro hematopoietic microenvironments. Meth MSCs were primarily cultured. The desmin protein was examined by immunohistochemical assay five days later. The MSCs were randomly divided into 8 groups, i. e., the control group, the supernatant from cultured bone marrow stroma cells group, 100,200,300μg/mL APS added in the DMEM/F12 medium experimental groups, and 100,200, 300 ug/mL APS intervened medium groups. The effects of the proliferation activities of MSCs were detected using MTT-Imethod. The c-kit protein of the MSCs was stained by immunohistochemistry. Results The desmin protein was positive in the isolated cultured MSCs. Results of MTF method showed the proliferation of MSCs in APS intervened medium groups was significant. The strong positive c-kit immunoreactivity existed in APS intervened medium groups. The strong positive c-kit immunoreactivity was present in the cytoplasmia of the MSCs in the DMEM/F12 medium experimental groups and the APS intervened medium groups. Cenclusions The APS intervened MSC medium could effectively change the growth properties of MSCs, obviously promote the proliferation of MSCs and c-kit expression. The c-kit protein might play some regulative roles in the proliferation of the MSCs.
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