不同组织来源间充质干细胞体外成骨分化能力的比较研究

发布时间：2018-03-11 15:30

本文选题：脂肪间充质干细胞　切入点：骨髓间充质干细胞　出处：《中国骨质疏松杂志》2017年05期 　论文类型：期刊论文

【摘要】：目的比较成人脂肪间充质干细胞(ASCs)、脐带间充质干细胞(UC-MSCs)和成人骨髓间充质干细胞(BMSCs)的成骨能力,选择优势干细胞种类作为应用于骨组织工程治疗骨缺损的种子细胞。方法采用含10%胎牛血清的DMEM/Ham’s F-12培养液培养3种MSCs。取3种MSCs的P3代,通过CCK8方法检测其增殖能力;通过流式细胞仪进行鉴定;通过碱性磷酸酶(ALP)和茜素红染色检测骨分化蛋白ALP的分泌和矿化钙结节的沉积,并对钙结节进行定量分析;通过实时荧光定量PCR(RT-q PCR)方法检测骨再生相关基因的表达。结果 3种P3代MSCs在3~5d之间增殖均处于对数生长期;流式鉴定3种细胞的表面标志物阳性率:CD44、CD90和CD105均高于97%,阴性率:CD14、CD34和CD45均低于1%;ALP染色结果显示3种MSCs成骨诱导9d时,细胞内均表达ALP,茜素红染色结果显示成骨诱导18d时,均呈现较好的矿化能力,BMSCs和UC-MSCs成骨诱导后形成的钙结节无显著性差异;RT-q PCR结果显示3种MSCs成骨诱导组相比较于对照组,成骨再生相关基因Osterix、ALP、I型胶原(COL1)和骨钙素(OCN)均显著性高表达;3种MSCs成骨诱导9d时,UC-MSCs实验组的COLI基因表达显著性高于BMSCs,成骨诱导18d时,ASCs实验组的Osterix基因表达显著性高于BMSCs。结论 ASCs和UC-MSCs具有一定的成骨矿化能力,有望成为骨组织工程治疗骨缺损的种子细胞。
[Abstract]:Objective to compare the osteogenic ability of adult adipose mesenchymal stem cells (ASCs), umbilical cord mesenchymal stem cells (UC-MSCs) and adult bone marrow mesenchymal stem cells (BMSCs). Methods three kinds of MSCs were cultured in DMEM/Ham's F-12 culture medium containing 10% fetal bovine serum. P3 generation of three kinds of MSCs were obtained and the proliferative ability was detected by CCK8 method. The secretion of bone differentiation protein (ALP) and the deposition of mineralized calcium nodules were detected by alkaline phosphatase (ALP) and alizarin red staining. The expression of bone regeneration related genes was detected by real-time fluorescence quantitative PCR(RT-q. Results the proliferation of three P3 generation MSCs was in logarithmic growth phase within 3days. The positive rates of CD105 and CD90 in three kinds of cells were higher than 97, and the negative rates of CD14, CD34 and CD45 were lower than those of 1T. The results showed that ALP was expressed in the three kinds of MSCs cells after 9 days of osteogenesis, and the results of alizarin red staining showed that at 18 days of osteogenesis. The results of RT-Q PCR showed that there was no significant difference between the three MSCs osteogenic groups and the control group. The expression of COLI gene in UC-MSCs group was significantly higher than that in BMSCs group on the 9th day after osteogenesis induction. The expression of Osterix gene in the experimental group was significantly higher than that in BMSCs group on the 18th day after osteogenesis induction, and the expression of Osterix gene in the experimental group was significantly higher than that in the control group at the 18th day after osteogenesis induction. Conclusion ASCs and UC-MSCs have some osteogenic mineralization ability. It is expected to be a seed cell for bone tissue engineering in the treatment of bone defect.
【作者单位】：天津市康婷生物工程有限公司;
【基金】：天津市自然科学基金(14JCYBJC43900)
【分类号】：R68

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