骨髓间充质干细胞体外诱导向内皮和上皮细胞的分化

背景：骨髓来源的间充质干细胞在体外具有多系分化潜能,但其在体外向肺组织细胞的分化能力尚存在争议。目的：体外诱导验证小鼠骨髓间充质干细胞向内皮细胞和上皮细胞分化的能力。方法：分离小鼠骨髓来源的间充质干细胞,以内皮诱导液向内皮细胞分化。另外将小鼠骨髓间充质干细胞分别在以下诱导培养基中进行上皮诱导3周：单纯上皮诱导培养液,上皮诱导培养液加10μg/L转化生长因子β1,并以未经诱导的小鼠骨髓间充质干细胞作为阴性对照,肺泡上皮作为阳性对照。结果与结论：小鼠骨髓间充质干细胞在上皮诱导培养液中诱导培养3周后,部分细胞由梭形变为典型的卵石样上皮细胞形态,诱导后约 60%细胞表达广谱上皮细胞标志pan-CK,RT-PCR结果显示分化后的细胞表达上皮细胞特异标志CK18,未经诱导的间充质干细胞未表达。小鼠骨髓间充质干细胞在内皮诱导24h后即出现了典型的血管网状结构,vWF免疫荧光染色显示约 70%的细胞呈阳性,RT-PCR 显示分化后的细胞表达内皮细胞特异性标志CD31、vWF和 CD34。提示骨髓来源的间充质干细胞体外诱导培养具有跨胚层多系分化能力。

Epithelial and endothelial differentiation of mouse bone marrow mesenchymal stem cells in vitro

BACKGROUND： Bone marrow mesenchymal stem cells （BMSCs） have multiple differentiation potential in vitro, but the in vitro differentiation of BMSCs to the lung tissue is still controversial. OBJECTIVE： To verify the ability of mouse BMSCs differentiating to endothelial cells and epithelial cells in vitro. METHODS： Mouse BMSCs were isolated and induced by endothelial-induced fluid to differentiate into endothelial cells. In addition, isolated mouse BMSCs were induced in these epithelial induction media for 3 weeks： a simple epithelial induction medium; epithelial induction medium plus 10 μg/L transforming growth factor β1. Non-induced mouse BMSCs were used as negative controls and the alveolar epithelium as a positive control. RESULTS AND CONCLUSION： After cultured in the epithelial induction medium for 3 weeks, some cells changed from the spindle cells into a typical pebble-like epithelial cell. 60% induced cells expressed epithelial cell marker pan-CK, and RT-PCR showed that differentiated epithelial cells expressed cell-specific signs CK18. There was no CK18 expression in non-induced mouse BMSCs. Mouse BMSCs in the epithelial induction medium for 24 hours, typical vascular network structure appeared. vWF immunofluorescence staining showed that about 70% of cells were positive, and RT-PCR showed that differentiated cells expressed CD31, vWF, and CD34. It is indicated that BMSCs has in vitro cross-mesodermal multilineage differentiation ability.