干细胞诱导、细胞系诱导和原代分离3种提取成骨细胞的方法比较

发布时间：2018-04-18 00:14

本文选题：骨髓 + 间质干细胞　；参考：《中国组织工程研究》2017年17期

【摘要】：背景:成骨细胞是骨组织构建的重要组织细胞之一。成骨细胞取材困难,不同方法获得的成骨细胞纯度及钙化能力不尽相同。目的:比较小鼠骨髓间充质干细胞成骨诱导、细胞系MC3T3-E1成骨诱导和小鼠颅骨原代细胞培养3种方法获得的成骨细胞的纯度及钙化能力。方法:采用贴壁纯化法分离出小鼠骨髓间充质干细胞,传至3代后用成骨诱导培养基诱导21 d;将前成骨细胞系MC3T3分别用成骨诱导培养液1和成骨诱导培养液2诱导21 d;使用Ⅰ型胶原酶消化法取得乳鼠颅骨原代成骨细胞。对3种方法获得的成骨细胞进行茜素红钙结节染色,观察其成骨特性。结果与结论:(1)骨髓间充质干细胞诱导后钙结节平均为16.3个/低倍镜视野;(2)前成骨细胞系MC3T3诱导培养液1组可观察到稀疏的钙结节,平均为1.7个/低倍镜视野,诱导培养液2组可观察到密集的钙结节,平均为44.6个/低倍镜视野,两种诱导液相比钙结节形成能力差异有显著性意义(P0.01);(3)原代成骨细胞钙结节平均为0.6个/低倍镜视野;(4)除MC3T3诱导液1组与原代成骨细胞相比差异无显著性意义外,其余两两相比成骨能力差异均有显著性意义(P0.01);(5)由于骨髓间充质干细胞分离培养具有更多不可控因素,因此MC3T3细胞用含有地塞米松的诱导培养液2进行成骨诱导方法较好。
[Abstract]:Background: osteoblasts are one of the important tissue cells in bone tissue construction.The purity and calcification ability of osteoblasts obtained by different methods are different.Aim: to compare the purity and calcification of mouse bone marrow mesenchymal stem cells (BMSCs) induced by osteogenic induction, MC3T3-E1 osteogenesis induction and primary culture of mouse skull cells.Methods: mouse bone marrow mesenchymal stem cells were isolated by adherent purification.The osteoblasts were induced by osteoblast induction medium for 21 days, MC3T3 cells were induced with osteogenic medium 1 and osteogenic culture medium 2 for 21 days, and the primary osteoblasts were obtained by type I collagenase digestion.The osteoblasts obtained by three methods were stained with alizarin red calcium nodule and their osteogenic characteristics were observed.Results and conclusion the average number of calcium nodules induced by bone marrow mesenchymal stem cells (BMSCs) was 16.3 / LVE2) the sparse calcium nodules were observed in the culture medium of preosteoblast cell line MC3T3, with an average of 1.7 / LVEL.Dense calcium nodules were observed in group 2, with an average of 44.6 / low power field of vision.There was significant difference in the ability of calcium nodule formation between the two inducers (P 0.01). The average calcium nodule of primary osteoblasts was 0.6 / low magnification field of vision (LVE4). There was no significant difference between group 1 and primary osteoblasts except MC3T3 inducer group 1 and primary osteoblast.There were significant differences in osteogenic ability between the other two groups. (P 0.01) because the isolation and culture of bone marrow mesenchymal stem cells had more uncontrollable factors, it was better for MC3T3 cells to use dexamethasone induced culture medium 2 to induce osteogenesis.
【作者单位】：华中科技大学同济医学院附属协和医院骨科;
【基金】：国家自然科学基金资助(81472134)~~
【分类号】：R318.08;R68

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