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目的研究间充质干细胞(MSC)对脐血CD34+细胞体外扩增能力和黏附分子表达的影响。方法从正常人骨髓中分离扩增MSC,通过免疫表型和向成骨细胞及脂肪细胞分化能力对其鉴定;将脐血CD34+细胞接种到MSC或其他培养液中,比较不同培养条件对造血细胞扩增能力、集落形成能力及黏附分子表达的影响。结果MSC表达Thy1、SH2、SB10、CD44、CD13、CD49e和CD29,不表达CD34、CD45、HLADR、CD14和CD31,经过诱导可以向成骨细胞和脂肪细胞分化;实验组在MSC和细胞因子作用下,扩增8d后有核细胞、CD34+、CD34+CD38-、CD34+CD62L+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表达较扩增前无明显变化,虽然ICAM1和L选择素表达下降,但实验组CD34+CD62L+和CD34+CD54+细胞的绝对数显著增加。结论MSC可为造血干细胞体外扩增提供适宜的微环境,有助于抑制HSC分化并保持其造血重建潜能和归巢能力。
Objective To investigate the effects of mesenchymal stem cells (MSCs) on the expansion of cord blood CD34 + cells in vitro and the expression of adhesion molecules. Methods MSCs were isolated from normal human bone marrow and identified by immunophenotyping and differentiation into osteoblasts and adipocytes. Cord blood CD34 + cells were seeded into MSC or other culture medium. The effects of different culture conditions on hematopoietic cells Amplification ability, colony formation ability and expression of adhesion molecules. Results MSC expressed Thy1, SH2, SB10, CD44, CD13, CD49e and CD29 but did not express CD34, CD45, HLADR, CD14 and CD31, and induced to differentiate into osteoblasts and adipocytes. MSCs and cytokines , And there were nucleated cells after 8 days of amplification. The numbers of CD34 +, CD34 + CD38-, CD34 + CD62L + cells and CFUCs were 145.57 ± 17.89, 37.47 ± 13.78, 69.78 ± 50.07, 10.74 ± 5.89 and 20.73 ± 5.54 times, respectively, The expression of ALCAM, VLAα4, VLAα5, VLAβ1, HCAM, PECAM and LFA1 in CD34 + cells did not change significantly after amplification. Although the expression of ICAM1 and L-selectin decreased, CD34 + CD62L + and CD34 + CD54 + The absolute number of cells is significantly increased. Conclusion MSC can provide a suitable microenvironment for the expansion of hematopoietic stem cells in vitro and help to inhibit HSC differentiation and maintain its hematopoietic reconstitution potential and homing ability.