1,25-(OH)2D3在促进大鼠外周血间充质干细胞扩增中的作用
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江苏省自然科学基金重点项目(BK2006729)和江苏省卫生厅兴卫工程医学重点人才项目(RC2007060)


Action of 1,25-(OH)2D3 in the expanding of rat peripheral blood mesenchymal stem cells
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    目的:探讨大鼠外周血中是否存在非黏附间充质干细胞(MSCs)以及1,25-(OH)2D3能否刺激它们形成成纤维细胞集落形成单位(CFU-f)-方法:从大鼠外周血中分离单核粒细胞(PBMGC)组份,在缺乏和添加10-8 mol/L 1,25-(OH)2D3条件下进行贴壁培养-培养至4天和8天时,将未贴壁细胞转移到新的培养皿中继续培养,即:“pour-off”培养,培养16天形成的细胞进行亚甲基蓝染色,计数CFU-f数目,并对4天“pour-off”培养形成的细胞进行免疫细胞化学染色-结果:PBMGC培养及4天和8天非黏附PBMGC培养都能形成CFU-fs,而1,25-(OH)2D3能够明显地促进它们形成CFU-f-非黏附细胞形成的克隆表达波形蛋白(vimentin)-α-平滑肌肌动蛋白(α-SMA)和Ⅲ型胶原,而不表达Ⅰ型胶原-结论:大鼠外周血中存在非黏附MSCs,在体外培养能够形成CFU-fs,而1,25-(OH)2D3具有促进外周血MSC扩增的作用-

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    Objective:To determine whether the non-adherent mesenchymal stem cells(MSCs) are present in rat peripheral blood and whether 1,25-(OH)2D3 can stimulate them to form colony forming unit-fibroblastic(CFU-f). Methods:Peripheral blood mononuclear and granular cells(PBMGC)fraction was isolated from the blood of six-week old male Wistar rat and were cultured in the absence or presence of 10-8 mol/L 1,25-(OH)2D3 for 16 days After 4 days and 8 days,the non-adherent cells were transferred into another fresh petri dishes to culture consensually until to day 16(“pour-off” culture). The resulting cells were stained with methylene blue and the number of colonies was counted. The resulting cells from 4-day “pour-off” cultures were strained immunocytochemically for vimentin,α-smooth muscle actin(α-SMA),type Ⅰ collagen and type Ⅲ collagen. Results:Not only total PBMGC,but also non-adherent PBMGC can give rise to CFU-fs and the treatment of 10-8 mol/L 1,25-(OH)2D3 increased their CFU-f forming efficacy significantly. The resulting cells from 4-day “pour-off” cultures expressed vimentin,α-SM actin and type Ⅲ collagen,but did not express type Ⅰ collagen. Conclusion:The non-adherent mesenchymal stem cells(MSCs) are present in rat peripheral blood and the treatment of 1,25-(OH)2D3 can stimulate them to form CFU-fs.

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刘 波,任永信,苗登顺,曹晓建.1,25-(OH)2D3在促进大鼠外周血间充质干细胞扩增中的作用[J].南京医科大学学报(自然科学版),2009,29(6):771-774

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  • 收稿日期:2008-12-08
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