肾组织干细胞向肾小管上皮细胞的诱导分化

摘要

目的：探讨肾组织干细胞向肾小管上皮细胞的诱导分化。方法分离肾乳头肾组织干细胞，对照组加入正常干细胞培养基，诱导组应用含activin A、BMP-7、维甲酸3种细胞因子的诱导培养基与肾上皮细胞培养基交替诱导，同时与缺氧损伤的肾小管上皮细胞共培养，通过细胞流式、免疫荧光、qRT-PCR的方法评估诱导效率。结果细胞流式结果提示，对照组肾组织干细胞CK-18阳性率为6.5%，诱导组CK-18阳性率为44.2%；免疫荧光结果提示，诱导组成熟上皮细胞标记CK-18、E-cadherin、ZO-1部分阳性表达；qRT-PCR结果提示，诱导组成熟上皮细胞标记E-cadherin、AQP-1基因表达水平较对照组显著升高（P<0.01）。结论肾组织干细胞在体外可以诱导分化为肾小管上皮细胞。%Objective To investigate the differentiation capability of kidney stem cells (KSCs) into renal tubular epithelial cells (RTECs). Methods KSCs isolated from the renal papilla of 4-week-old SD rats were co-cultured with hypoxia-exposed RTEC in induced medium (containing activin A, BMP-7, and retinoic acid) and renal epithelial cell growth medium (REGM) alternately. The KSCs cultured in MSC medium served as the control. The KSC differentiation rates in both groups were determined using flow cytometry, immunofluorescence assay and qRT-PCR. Results Flow cytometry showed a CK-18 positive rate of 6.5%in the control KSC group and of 44.2%in the induced group. Immunofluorescence assay detected the positivity for mature epithelial cell markers CK-18, E-cadherin, and ZO-1 in the induced cells. The results of qRT-PCR showed significantly increased expression of E-cadherin and AQP-1 mRNAs in the induced cells compared with the control cells (P<0.01). Conclusion Rat KSCs can be induced to differentiate into RTECs in vitro.