小鼠骨髓间充质干细胞培养及鉴定

何琳琳

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小鼠骨髓间充质干细胞培养及鉴定

首发时间：2019-07-11

何琳琳 ^{1
2}
何琳琳（1972-），女，硕导，主要研究方向：天然活性产物的活性评价及作用机理研究

1、陕西理工大学生物科学与工程学院，陕西汉中 723000
2、陕西理工大学维生素D生理与应用研究所，陕西汉中 723000

摘要：[目的] 从C57BL/6小鼠骨髓培养骨髓间充质干细胞（bone marrow-derived mesenchymal stem cells, BM-MSCs）并鉴定其成骨和成脂分化潜能。[方法] 以全骨髓贴壁法培养小鼠BM-MSCs。为鉴定小鼠BM-MSCs成骨分化潜能，将其成骨诱导培养后，进行碱性磷酸酶（alkaline phosphatase, ALP）染色和茜素红（alizarin red S, ARS）染色，实时定量PCR（real-time quantitative polymerase chain reaction, Real-time qPCR）检测成骨分化相关基因OCN和ALP的mRNA表达变化；为鉴定其成脂分化潜能，将其经成脂诱导和维持培养后，进行油红O染色，Real-time qPCR检测成脂分化相关基因PPARγ2的mRNA表达变化。[结果] 从C57BL/6小鼠骨髓成功地培养得到小鼠BM-MSCs。成骨诱导培养后有ALP阳性细胞和矿化骨节形成，OCN和ALP的mRNA表达极显著上升；成脂诱导和维持培养后有脂滴形成，PPARγ2的mRNA表达极显著上升。[结论] 从C57BL/6小鼠骨髓中培养得到具有成骨和成脂分化潜能的小鼠 BM-MSCs，为将其作为种子细胞应用于临床研究或基础研究鉴定基础。

关键词：骨髓间充质干细胞; 鉴定; 分化潜能

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Culture and Identification of Mouse Bone Marrow-Derived Mesenchymal Stem Cells

HE Linlin ^{1
2}
何琳琳（1972-），女，硕导，主要研究方向：天然活性产物的活性评价及作用机理研究

1、School of Biological Science and Engineering, Shaanxi University of Technology, Hanzhong 723000, China
2、 Vitamin D Research Institute, Shaanxi University of Technology, Hanzhong 723000, China

Abstract：[Objective] Bone marrow-derived mesenchymal stem cells (BM-MSCs) were cultured from C57BL/6 mouse bone marrow, and their osteogenic and adipogenic differentiation potential were tested. [Methods] Mouse BM-MSCs were cultured by using the whole bone marrow adherence method. To identify the potential of their osteogenic differentiation, alkaline phosphatase staining and Alizarin Red S staining were tested after osteogenic induction culture, and reverse transcription polymerase chain reaction (RT-PCR) was used to analyze mRNA expression of the osteogenic differentiation related genes OCN and ALP. To identify the potential of their adipogenic differentiation, oil red O staining was tested after adipogenic induction and maintenance culture, and RT-PCR was used to analyze mRNA expression of PPARγ2. [Results] BM-MSCs were successfully cultured from bone marrow of C57BL/6 mice. ALP positive cells and mineralized bone nodules were showed up after osteogenic induction, and mRNA expression of OCN and ALP increased very significantly. Lipid drops were formed after adipogenic induction and maintenance culture, and mRNA expression of PPARγ2 increased very significantly. [Conclusion] BM-MSCs with the potential of osteogenic and adipogenic differentiation were obtained from mouse bone marrow, which will provide the foundation for the clinical and basic research using mouse BM-MSCs as seeding cells.

Keywords： bone marrow-derived mesenchymal stem cells (BM-MSCs) identification differentiation potential

基金：

1. 陕西省自然科学基础研究计划项目（2018JM3033）

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何琳琳. 小鼠骨髓间充质干细胞培养及鉴定[EB/OL]. 北京：中国科技论文在线 [2019-07-11]. https://www.paper.edu.cn/releasepaper/content/201907-22.

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论文编号	201907-22
论文题目	小鼠骨髓间充质干细胞培养及鉴定
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