小鼠CD45-CD31+肺侧群细胞诱导分化的研究

Differentiation of mouse CD45-/CD31+ lung side population cells into endothelial cells in vitro

摘要:

摘要：目的  探讨在体外诱导分化肺侧群细胞（SP）的特性。方法  取小鼠肺组织，流式细胞术分选小鼠白细胞分化抗原（CD）45-/CD31+肺SP；免疫荧光检测分选肺SP三磷酸腺苷结合转运蛋白G超家族成员2（ABCG2）的表达；逆转录聚合酶链反应（RT-PCR）检测肺SP中ABCG2、酪氨酸激酶2（Tie2）及血管性血友病因子（vWF）的表达。细胞体外分化诱导14 d后，免疫荧光检测细胞vWF的表达；RT-PCR检测分化诱导前后细胞中ABCG2和vWF的表达。结果  流式细胞术成功分选出CD45-/CD31+肺SP细胞，免疫荧光检测其表达ABCG2；RT-PCR检测SP表达ABCG2和Tie2，不表达vWF。主群细胞表达vWF；分化诱导前的肺SP表达ABCG2；分化诱导后的肺SP表达vWF。结论  CD45-/CD31+肺SP是血管内皮细胞的祖细胞，具有干细胞分化特性。在体外培养可分化为血管内皮细胞。

Abstract:

Abstract: Objective To investigate the characteristics of differentiation of lung side population (LSP) cells in vitro. Methods Mouse lung tissue was taken, and mouse CD45-/CD31+ LSP cells were sorted by flow cyto-metry. Immunofluorescence was used to detect the expression of ATP-binding cassette sub-family G member 2 (ABCG2) in the isolated LSP cells. RT-PCR was applied to detect the expression of ABCG2, Tie2 and vWF in mouse CD45-/CD31+ LSP cells. After the LSP cells were cultured for 14 days, the expression of vWF was detected by immunofluorescence, and the mRNA expression of ABCG2 as well as vWF of both freshly isolated LSP cells and cultured LSP cells were examined by RT-PCR. Results Flow cytometry sorted out the CD45-/CD31+ LSP cells, and immunofluorescence tested that they expressed ABCG2. RT-PCR results revealed that LSP cells expressed ABCG2 as well as SMA and Tie2, but did not express vWF. LMP highly expressed vWF. The CD45-/CD31+ LSP cells expressed ABCG2, but did not express vWF before induction of differen-tiation; while the cells expressed vWF in stead of ABCG2 after the induction of differentiation. Conclusions CD45-/CD31+ LSP cells might be the progenitor cells of vascular endothelial cells, which possess the characteristics of stem cell differentiation, and can be differentiated into vascular endothelial cells in vitro.