无菌回收膝关节镜术后引流液体外培养自体间充质干细胞的可行性

发布时间：2019-03-07 10:36

【摘要】：背景:目前间充质干细胞在骨科中的应用前景极为广阔,尤其是对于治疗关节软骨缺损疾病表现出巨大的潜力,骨髓是间充质干细胞的主要来源,但常规髂骨穿刺法抽取骨髓,来源有限,技术要求严格,限制了其应用程度,因此,探寻新的方便有效的细胞来源具有重要意义。目的:探讨膝关节镜术后引流液体外培养自体间充质干细胞的可行性。方法:选取行膝关节镜手术患者8例,在关闭切口前置特制的无菌血袋,收集术后引流液,采用羟乙基淀粉沉淀法及密度梯度离心法分离培养间充质干细胞,观察细胞形态,绘制生长曲线,流式细胞仪鉴定表面标志物,进行三系诱导分化染色鉴定。结果与结论:(1)细胞呈梭形,贴壁生长,细胞增殖能力好,表面表达CD44、CD90、CD105、CD73,而不表达CD45;(2)通过标准方法诱导,间充质干细胞能向成骨细胞、脂肪细胞及软骨细胞分化,表明培养的细胞为间充质干细胞;(3)8例样本均培养出间充质干细胞,其细胞形态、增殖能力、细胞表型等无明显差异;(4)该结果表明,膝关节镜术后引流液可以培养出间充质干细胞,且细胞质量稳定。
[Abstract]:Background: at present, the application of mesenchymal stem cells in orthopedics is very broad, especially for the treatment of articular cartilage defects. Bone marrow is the main source of mesenchymal stem cells. The limited sources and strict technical requirements limit its application. Therefore, it is of great significance to explore new convenient and effective cell sources. Objective: to investigate the feasibility of autologous mesenchymal stem cells (MSCs) cultured in vitro after knee arthroscopy. Methods: eight patients undergoing knee arthroscopic surgery were selected to collect the postoperative drainage fluid and culture mesenchymal stem cells (MSCs) by hydroxyethyl starch precipitation and density gradient centrifugation to observe the morphology of mesenchymal stem cells (MSCs). The growth curve was drawn, the surface markers were identified by flow cytometry, and the three-line induced differentiation staining was carried out. Results & conclusion: (1) the cells were spindle-shaped, adherent to the wall, and had good proliferation ability. The cells expressed CD44,CD90,CD105,CD73, on the surface but did not express CD45;. (2) the mesenchymal stem cells could differentiate into osteoblasts, adipocytes and chondrocytes by standard method, indicating that the cultured cells were mesenchymal stem cells. (3) there was no significant difference in cell morphology, proliferation ability and phenotype of mesenchymal stem cells (MSCs) in all 8 samples. (4) the results showed that mesenchymal stem cells could be cultured in the drainage solution after knee arthroscopy, and the cell quality was stable.
【作者单位】：山东中医药大学;山东中医药大学附属医院;
【基金】：济南赛尔生物科技股份有限公司
【分类号】：R687.4

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