体外培养骨髓干细胞及趋化因子受体CXCR4的表达

Bone marrow mesenchymal stem cells cultured in vitro and chemokine receptor CXCR4 expression

背景:有实验证明骨髓间充质干细胞表面CXCR4的表达水平决定了细胞向靶组织归巢及随后血管生成的效率。目的:分析骨髓间充质干细胞的分离、培养、鉴定及多向分化潜能,探讨细胞表面CXCR4的表达在促血管再生作用领域的意义。方法:无菌条件下骨穿针垂直穿刺髂后上棘处抽取猪骨髓,并行分离及培养,得到传代的骨髓间充质干细胞,并行体外细胞凋亡检测,骨髓间充质干细胞表面蛋白CXCR4的检测:并行骨髓间充质干细胞的凋亡周期检测。结果与结论:实验分离骨髓间充质干细胞的细胞形态比较均一。传代培养后,细胞不再以集落方式生长,而呈分布均匀的纺锤形。细胞在4d左右开始快速增长,1周后细胞生长进入稳定期。流式细胞仪检测发现细胞表面CXCR4的表达较多。证明实验成功分离骨髓干细胞,CXCR4蛋白在细胞体外培养中明显高表达,提高CXCR4的表达将为骨髓间充质干细胞移植促进血管再生、治疗下肢缺血性疾病提供临床治疗依据。

BACKGROUND：Experiments have demonstrated that the expression level of CXCR4 on bone marrow mesenchymal stem cel s determines cel homing to targeted tissue and subsequent angiogenesis. OBJECTIVE：To analyze the isolation, culture, identification and multi-differentiation potential of bone marrow mesenchymal stem cel s and investigate the significance of chemokine receptor CXCR4 expression on bone marrow mesenchymal stem cel s in promotion of angiogenesis. METHODS：Under sterile condition, bone marrow mesenchymal stem cel s were isolated, cultured and purified by the while bone marrow adherence method. Cel apoptosis in vitro, CXCR4 protein expression on the bone marrow mesenchymal stem cel s were detected, and cel cycle was analyzed. RESULTS AND CONCLUSION：The isolated bone marrow mesenchymal stem cel s exhibited uniform morphology. After passage, cel s were spindle-shaped. Cel s began to grow rapidly at about 4 days and grew stably 1 week later. Flow cytometry results showed increased CXCR4 expression on bone marrow mesenchymal stem cel s. These findings suggest that bone marrow mesenchymal stem cel s were successful y isolated, CXCR4 protein was highly expressed on cel surface after in vitro culture, and increased CXCR4 expression wil provide theoretical evidence for bone marrow mesenchymal stem cel transplantation in promotion of angiogenesis and treatment of lower limb ischemic diseases in the clinic.