| 李强强,谢亚东,张怀斌,杨国清,梁文强,王勇平.SD大鼠骨髓间充质干细胞分离培养及鉴定的实验研究[J].现代生物医学进展英文版,2021,(13):2410-2414. |
| SD大鼠骨髓间充质干细胞分离培养及鉴定的实验研究 |
| Isolation, Cultivation and Identification of SD Rat Bone Marrow Derived Mesenchymal Stem Cells |
| Received:January 28, 2021 Revised:February 23, 2021 |
| DOI：10.13241/j.cnki.pmb.2021.13.003 |
| 中文关键词: 骨髓间充质干细胞 SD大鼠 全骨髓贴壁法 流式细胞仪 |
| 英文关键词: Bone marrow mesenchymal stem cells SD rats Whole bone marrow adherent method Flow cytometry |
| 基金项目:国家自然科学基金项目（81960398）；甘肃省自然科学基金项目（20JR5RA369）；兰州市城关区科技计划项目（No.2018-1-2） |
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| 中文摘要: |
| 摘要 目的：探讨应用全骨髓贴壁法体外分离培养SD大鼠骨髓间充质干细胞（BMSCs)的可行性，研究其生物学特性，为骨组织工程提供种子细胞。方法：取SPF级5周龄健康SD大鼠2只，脱颈处死,分离双下肢股骨、胫骨，全骨髓贴壁法分离培养、纯化BMSCs；通过倒置显微镜观察原代、传代细胞生长情况、绘制生长、贴壁率曲线，研究其生物学特性；流式细胞仪检测表面标志物、诱导成成骨等方法进行鉴定。结果：应用全骨髓贴壁法可在体外分离出活性好、纯度高的BMSCs。倒置显微镜下可见原代细胞呈梭形、多角形，传代细胞形态均一呈纤维样；P3代BMSCs经流式细胞鉴定：CD44、CD90高表达，CD31、CD45低表达；定向诱导向成骨细胞分化，可见明显矿化结节。结论：证实应用全骨髓贴壁培养法体外可成功分离BMSCs，所分离培养、纯化的细胞生物学稳定，纯度高、活性好，具有多向分化潜能，能为骨组织工程、骨质疏松症和骨折不愈合疾病的研究提供种子细胞。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the feasibility of the whole bone marrow attachment method to isolate and culture SD rat bone marrow mesenchymal stem cells in vitro, study their biological characteristics, and provide seed cells for bone tissue engineering. Methods: Two healthy SD rats aged 5 weeks with SPF grade were sacrificed by neck dissection. Femur and tibia of lower limbs were isolated, and BMSCs were cultured and purified by whole bone marrow adherent method. The growth of primary and passage cells was observed by inverted microscope, the growth and adherent rate curves were plotted to study their biological characteristics, and the surface markers, fat induction and bone formation were identified by flow cytometry. Results: BMSCs with good activity and high purity could be isolated by whole bone marrow adherent method in vitro.Under inverted microscope, the primary cells showed long spindle shape, short spindle shape and polygon shape, and the morphology of the passage cells was uniform and fibro-like.BMSCs were identified by flow cytometry: CD44 and CD90 were highly expressed, while CD31 and CD45 were low. The differentiation of osteoblasts was induced, and mineralized nodules were visible. Conclusion: BMSCs can be successfully isolated by whole bone marrow adherent culture method in vitro. The isolated, cultured and purified BMSCs are biologically stable, with high purity, good activity and multidirectional differentiation potential, which can provide seed cells for bone tissue engineering research. |
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