摘要
目的分离培养和鉴定大鼠骨髓间充质干细胞(BMSCs),探讨高浓度葡萄糖对BMSCs增殖及凋亡的影响。方法贴壁培养法分离纯化Wister大鼠BMSCs,体外培养和连续传代。将传至第三代的BMSCs用低糖-DMEM培养液(含葡萄糖5.6 mmol/L)制成细胞悬液,计数并接种于培养瓶中,待细胞贴壁后,分组加入含不同浓度葡萄糖(5.6 mmol/L、11 mmol/L、25 mmol/L、40mmol/L)的DMEM培养液,短期组培养3～4天,长期组培养4周。分别收集细胞,噻唑蓝(MTT)法检测细胞增殖,Annexin-V/PI法检测其凋亡。结果在实验研究中,与高糖对内皮祖细胞(endothelial p rogenitor cells,EPCs)等的影响不同,一定浓度的高糖并不显著抑制BMSCs的增殖,并不促进BMSCs的凋亡率显著增加。25 mmol/L的高糖有助于BMSCs成骨分化。结论短期和长期(4周)培养,BMSCs具有显著的高糖耐受性。与低糖条件相比,高糖提高了细胞的成骨分化能力。
Objective To culture and identify rat bone marrow mesenchymal stem cells （BMSCs） and To explore proliferation and apoptosis of BMSCs in HG-DMEM. Methods BMSCs were isolated and purified by the attachment culturemethod. Using low glucose culturing liquid made the third pericytes cells into suspension cell, counted cells and planted them in culture-bottles, then made DMEM culturing liquid of different glucose （5.6、 11、 25、 40mmol/L） Cultured. after 3 - 4days and 4weeks in vitro, we collected cells and studied the effect of high glucose for them.MTT assayswere used to detect the proliferative rates. Annexin-V/PI stains were used to detect the apoptotic rates. Results BMSCs proliferation in studies was not decreased by HG. apoptosis was not increased by HG in BMSC. BMSC treatment with HG （25 mM） favored osteogenic differentiation. Gonclusion During short-and long-term exposure up to 4 weeks BMSC display remarkable resistance against HG. HG （25 mM） in culture even significantly enhance osteogenic differentiation in vitro when compared to Low glucose （ 5.6 mM ） .
出处
《中国医药技术经济与管理》
2008年第9期62-68,共7页
China Pharmaceutical Technology Economics & Management
关键词
骨髓间充质干细胞
葡萄糖
增殖
凋亡
Mesenchymal stem cells
Glucose
proliferation
apoptosis
