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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。
Objective To investigate the potential of hair follicle culture supernatant to induce the lateral differentiation of rat bone marrow mesenchymal stem cells (MSCs) into hair follicle stem cells in vitro. Methods Rat MSCs were isolated and cultured completely by adherent method. MSCs of passage 3 were harvested and CD44 and CD29 were identified by immunocytochemical staining. The conditioned medium of hair follicles was used as the conditioned medium to induce MSCs. The morphological changes of the cells were observed under inverted phase contrast microscope. The induced cells were analyzed by immunocytochemistry and immunofluorescence cytochemistry with immunocytochemistry, reverse transcription - polymerase chain reaction (RT-PCR) to further identify the induced cytokeratin 15 expression. Results MSCs cultured in vitro showed positive expression of CD44 and CD29. After induction by the hair follicle supernatant, immunocytochemistry and immunofluorescence staining showed that some cytokeratin 15 was positive. Meanwhile, the expression of keratin 15 mRNA was also detected by RT-PCR. Conclusion Bone marrow-derived mesenchymal stem cells cultured in vitro can differentiate into hair follicle stem cell-like cells induced by hair follicle culture supernatant.