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目的:探讨脐血源性间充质干细胞(mesenchymal stem cells,MSCs)经成骨诱导分化后,其对淋巴细胞的免疫调节作用。方法:从人脐血中分离、培养及扩增MSCs,流式细胞仪测定细胞表型;将获得的MSCs在成骨诱导体系中诱导分化并鉴定,将诱导后的成骨细胞分别以不同剂量加入到外周血混合淋巴细胞培养体系和植物血凝素(PHA)刺激的外周血淋巴细胞转化体系中,用3H-TdR标记β液体闪烁计数仪检测细胞增殖,观察对混合淋巴细胞反应和淋巴细胞转化的影响。结果:从人脐血中分离获得的MSCs,呈成纤维样的细胞形态,CD29和CD105表达阳性,CD34和CD45表达阴性;在成骨诱导剂诱导下,具有成骨细胞样表型,可表达碱性磷酸酶,并形成矿化结节;人脐血源性MSCs经成骨诱导分化后在体外对混合淋巴细胞反应和PHA诱导的淋巴细胞转化均具有明显的抑制作用,抑制作用与细胞数量呈正相关。结论:脐血源性MSCs诱导为成骨细胞后对同种异体淋巴细胞具有免疫调节作用,为其作为骨组织工程异基因种子细胞来源打下基础。
Objective: To investigate the immunomodulatory effect of mesenchymal stem cells (MSCs) on lymphocytes after osteogenic differentiation. METHODS: MSCs were isolated, cultured and expanded from human umbilical cord blood. Cell phenotypes were determined by flow cytometry. MSCs were induced to differentiate and identified in osteogenic induction system. The induced osteoblasts were cultured in different doses Was added to the peripheral blood lymphocyte culture system and phytohemagglutinin (PHA) -stimulated peripheral blood lymphocyte transformation system, 3H-TdR labeled β liquid scintillation counter was used to detect cell proliferation, observed mixed lymphocyte reaction and lymphocyte The impact of conversion. RESULTS: MSCs isolated from human umbilical cord blood were fibroblast-like, with positive expression of CD29 and CD105, negative expression of CD34 and CD45, osteoblast-like phenotype induced by osteoinductive agent Alkaline phosphatase, and form mineralized nodules; human umbilical cord blood-derived MSCs after osteogenic differentiation in vitro mixed lymphocyte reaction and PHA-induced lymphocyte transformation were significantly inhibited, inhibition and cell number Was positively correlated. CONCLUSION: The cord blood-derived MSCs have the immunomodulatory effect on allogeneic lymphocytes induced by osteoblasts and lay a foundation for their origin as allogeneic seed cells for bone tissue engineering.