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目的:探讨骨髓间充质干细胞(MSC)的黏附分子表达及其对脐血造血干/祖细胞体外增殖的支持作用。方法:取正常骨髓单个核细胞贴壁培养,梭形细胞完全融合后传代,用流式细胞仪检测免疫表型;将脐血CD34+细胞接种到MSC或其他培养液上,比较不同培养条件对造血细胞扩增能力及黏附分子表达的影响。结果:MSC在体外培养中分为成纤维样成熟MSC(mMSC)和体积很小的圆形RS(rapidlyselfrenewingcells)细胞;两群细胞的CD34、CD45、CD3、CD19、CD33、HLADR、CD38均为阴性,而CD90、CD105、CD166、CD29、CD44、CD49e、CD54、CD13呈阳性,但RS细胞表面抗原表达的阳性率和平均荧光强度都明显低于mMSC,而CD117的表达高于mMSC;脐血CD34+细胞在MSC和细胞因子作用下,扩增8d后有核细胞、CD34+、CD34+CD38-细胞和CFUCs分别扩增(145.57±17.89)、(37.47±13.78)、(69.78±50.07)、(10.74±5.89)和(20.73±5.54)倍,均显著高于对照组,扩增后CD34+细胞的ALCAM、VLAα4、VLAα5、VLAβ1、HCAM、PECAM和LFA1表达较扩增前无显著变化。结论:MSC是一个异质性的细胞群体,可为造血干细胞(HSC)体外扩增提供适宜的微环境,有助于抑制HSC分化并保持其归巢能力。
Objective: To investigate the expression of adhesion molecules in bone marrow mesenchymal stem cells (MSCs) and its role in the proliferation of cord blood hematopoietic stem / progenitor cells in vitro. METHODS: Normal bone marrow mononuclear cells were adherently cultured, spindle cells were completely fused and passaged, and the immunophenotypes were detected by flow cytometry. Cord blood CD34 + cells were inoculated into MSC or other culture medium. The effects of different culture conditions on hematopoietic Cell Apoptosis and Adhesion Molecule Expression. Results: MSCs were divided into fibroblast-like mature MSC (mMSC) and very rapid circular regenerative cells (RSs) in vitro. The two groups of cells were negative for CD34, CD45, CD3, CD19, CD33, HLADR and CD38 , While the positive rate and average fluorescence intensity of CD90, CD105, CD166, CD29, CD44, CD49e, CD54 and CD13 were all significantly lower than that of mMSC, while the expression of CD117 was higher than that of mMSC; After 8 days of expansion, the number of CD34 +, CD34 + CD38- cells and CFUCs of the cells were expanded (145.57 ± 17.89), (37.47 ± 13.78), (69.78 ± 50.07) and (10.74 ± 5.89) and (20.73 ± 5.54) folds, which were significantly higher than that of the control group. The expression of ALCAM, VLAα4, VLAα5, VLAβ1, HCAM, PECAM and LFA1 in CD34 + cells did not change significantly before amplification. CONCLUSIONS: MSC is a heterogeneous cell population that provides a suitable microenvironment for the in vitro expansion of hematopoietic stem cells (HSCs) and helps to inhibit HSC differentiation and maintain its homing potential.