毛囊培养上清液诱导大鼠骨髓间充质干细胞分化的研究

目的探讨毛囊培养上清液诱导体外培养的大鼠骨髓间充质干细胞(MSCs)向毛囊干细胞横向分化的潜能。方法采用完全贴壁法分离培养大鼠MSCs,取第3代MSCs,用免疫细胞化学染色技术鉴定CD44和CD29。用毛囊培养上清液为条件培养液诱导MSCs,倒置相差显微镜观察细胞形态学变化,对诱导后的细胞进行角蛋白15的免疫细胞化学染色和免疫荧光细胞化学染色,逆转录-聚合酶链反应(RT-PCR)进一步鉴定诱导后细胞角蛋白15的表达。结果体外培养的MSCs,CD44、CD29表达阳性。经毛囊上清液诱导后,免疫细胞化学、免疫荧光细胞化学染色鉴定,部分细胞角蛋白15表达阳性;同时RT-PCR也检测到keratin 15 mRNA的表达。结论体外培养的骨髓间充质干细胞经毛囊培养上清液诱导可以分化为毛囊干细胞样的细胞。

THE DIFFERENTIATION OF MESENCHYMAL STEM CELLS INDUCED BY THE SUPERNATANT OF CULTURED HAIR FOLLICLE

Objective To observe the possibility of inducing mesenchymal stem cells(MSCs)to differetiate into hair follicle stem cells by the supernatant of cultured hair follicle EM>in vitro/EM> and to investigate the transdifferentiation potentiality of MSCs. Methods MSCs were isolated and cultured from rat bone marrow by complete adherence. MSCs of passage 3 were characterized with markers CD44 and CD29 by immuohistochemical staining technique. The stem cells were induced by the supernatant of cultured hair follicle. The morphological character was observed by inverse phase contrast microscopy. The expression of keratin 15 was detected by immunohistochemical staining technique and immunofluorescence staining technique. RT-PCR was further used to detect the expression of keratin 15. Results The isolated and separated MSCs were immunostaining positive in CD44 and CD29. After induced by the hair follicle conditioned medium EM>in vitro/EM>, MSCS could be partially identified by the positive staining for keratin 15, a specific antibody of hair follicle stem cells. After 3 weeks′ induction, keratin 15 was detected by RT-PCR in MSCs induced by supernatant of cultured hair follicle. Conclusion MSCs have the potential to differ