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目的:探讨脐带间充质干细胞(UC-MSC)体外与造血干细胞共培养后Notch信号分子的改变。方法:通过胶原酶消化方法分离UC-MSC,通过流式细胞仪检测以及成脂、成骨和成软骨诱导鉴定UC-MSC具备间充质干细胞的特性。进而,将UC-MSC与脐血CD34+造血干细胞(HSC)体外培养,实时PCR方法检测MSC及CD34+细胞表面Notch配体及受体表达以及表达是否存在变化;在共培养体系中加入Notch信号阻滞剂DAPT(γ-secretase抑制剂),比较Hes-1基因活化状态的改变。结果:体外实验显示:UC-MSC在形态学、细胞表面表型和诱导分化能力上均具备间充质干细胞的特性。UC-MSC及CD34+细胞表面存在Notch信号配体及受体的表达,共培养后Jagged 1、Notch1基因表达明显增加;共培养后CD34+细胞中的Hes-1基因表达明显增加而加入DAPT后Hes-1基因表达未检出明显改变。结论:UC-MSC支持造血中,Notch信号可能发挥重要作用。
Objective: To investigate the changes of Notch signaling molecules after co-culture of umbilical cord mesenchymal stem cells (UC-MSCs) with hematopoietic stem cells. Methods: UC-MSCs were isolated by collagenase digestion. UC-MSCs were identified as mesenchymal stem cells by flow cytometry and adipogenic, osteogenic and chondrogenic induction. Furthermore, UC-MSCs and cord blood CD34 + hematopoietic stem cells (HSCs) were cultured in vitro. Real-time PCR was used to detect the expression and expression of Notch ligand and receptor on the surface of MSC and CD34 + cells. Notch signaling was added to the co-culture system DAPT, a gamma-secretase inhibitor, was used to compare changes in the activation state of the Hes-1 gene. RESULTS: In vitro experiments showed that UC-MSC possesses the characteristics of mesenchymal stem cells in morphological, cell surface phenotype and differentiation-inducing ability. The expression of Notch signaling ligand and receptor existed on the surface of UC-MSC and CD34 + cells. The co-cultured Jagged 1 and Notch1 gene expressions were significantly increased. The expression of Hes-1 gene in CD34 + cells after co-culture increased significantly. 1 gene expression was not detected significant change. Conclusion: Notch signaling may play an important role in the hematopoiesis of UC-MSCs.