人骨骼肌来源不同表面标志物间充质干细胞特定亚群的多向分化潜能

发布时间：2018-04-22 16:33

本文选题：干细胞 + 组织工程　；参考：《中国组织工程研究》2017年13期

【摘要】：背景:骼肌来源的间充质干细胞,在骨形态发生蛋白4等因子的调节下具有多向分化潜能,但具体细胞亚群的分化情况尚不十分明确。目的:观察人骨骼肌来源的不同表面标志物的间充质干细胞特定亚群的多向分化潜能。方法:应用流式细胞分选技术,分选出具有不同细胞表面标志物的人骨骼肌干细胞亚群,根据CD56及CD34/CD144的表达差异,将细胞分成CD56~+,CD56~+CD34~+CD144~+,CD34~+CD144~+及未分选组共4组,对比不同细胞亚群组在骨形态发生蛋白4成骨诱导或骨形态发生蛋白4~+转化生长因子β3成软骨诱导后的差异。通过碱性磷酸酶染色后碱性磷酸酶阳性细胞的百分比评估细胞骨代谢情况。将各亚群细胞进行3D培养和细胞因子处理后,通过硝酸银染色评估其成骨分化程度,通过阿尔新蓝染色评价成软骨分化程度。结果与结论:(1)细胞骨代谢:经骨形态发生蛋白4处理后,4组细胞的碱性磷酸酶阳性细胞的百分比差异无显著性意义(P0.05);(2)细胞分化:不同表面标记物的4组细胞成骨及成软骨分化有明显差异,在相同成骨培养条件下,CD56~+组成骨能力最强;相同成软骨培养条件下,CD56~+,CD56~+CD34~+CD144~+组呈阿尔新蓝染色阳性,CD56~+CD34~+CD144~+组的染色更深且区域更大;(3)结果证实,成骨与成软骨潜能与不同的人骨骼肌间充质干细胞类型有关。人骨骼肌来源的间充质干细胞CD56~+亚群和成骨潜能有密切关系。同时CD56~+CD34~+CD144~+亚群分化能力更强,具有更强的成软骨潜能。
[Abstract]:Background: mesenchymal stem cells (MSCs) derived from the iliac muscle have the potential to differentiate in many ways under the regulation of bone morphogenetic protein 4 (BMP 4), but the differentiation of specific cell subsets is not very clear. Aim: to observe the multidirectional differentiation potential of mesenchymal stem cells (MSCs) with different surface markers derived from human skeletal muscle. Methods: the subsets of human skeletal muscle stem cells with different cell surface markers were selected by flow cytometry. According to the expression of CD56 and CD34/CD144, the cells were divided into four groups: CD56 ~ + CD56 ~ CD34 ~ CD144 ~ + CD34 ~ CD144~ and no group (n = 4). The difference of osteogenesis induced by bone morphogenetic protein 4 (BMP 4) or osteogenesis induced by transforming growth factor 尾 3 (TGF- 尾 3) was compared between different cell subgroups. Bone metabolism was evaluated by percentage of alkaline phosphatase positive cells after alkaline phosphatase staining. After 3D culture and cytokine treatment, the osteogenic differentiation was evaluated by silver nitrate staining and cartilage differentiation was evaluated by new blue staining. Results and conclusion: bone metabolism: the percentage of alkaline phosphatase positive cells in the 4 groups treated with bone morphogenetic protein 4 had no significant difference (P 0.05); osteoblasts of 4 groups with different surface markers were differentiated. And chondrogenic differentiation, In the same osteogenic culture condition, the ability of CD56 ~ + to form bone was the strongest, and in the same condition of chondrogenic culture, the positive staining of CD56 ~ + CD56 ~ CD34 ~ CD144 ~ ~ ~ + was found to be deeper and the area was larger than that of the control group (P < 0.05). Osteogenic and chondrogenic potential are associated with different types of human skeletal mesenchymal stem cells. CD56 ~ subsets of mesenchymal stem cells derived from human skeletal muscle are closely related to osteogenic potential. At the same time, CD56 ~ CD34 ~ CD144- subgroup differentiation ability is stronger, have stronger cartilage forming potential.
【作者单位】：郑州大学第一附属医院骨科;
【基金】：国家自然科学基金(81472136)~~
【分类号】：R318.08;R68

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