牙囊细胞条件培养液在大鼠脂肪间充质干细胞向成牙骨质分化中的作用

Effect of Dental Follicle Cell Conditioned Media on Cementoblast Differentiation of Adipose-derived Stem Cells

目的:探讨牙囊细胞条件培养液(DFCCM)在诱导大鼠脂肪间充质干细胞(ADSCs)向成牙骨质样细胞分化中的作用。方法:分离培养大鼠脂肪间充质干细胞、牙囊细胞,制备DFCCM。用DFCCM诱导ADSCs,通过MTT检测ADSCs增殖能力的变化;免疫荧光及实时定量PCR方法检测成骨关键蛋白-骨钙素(OCN)、碱性磷酸酶(ALP)及成牙骨质关键蛋白-牙骨质附着蛋白(CAP),牙骨质蛋白(CP23)的情况。结果:牙囊细胞条件培养液诱导ADSCs后可抑制ADSCs的增殖能力。DFCCM诱导7d后,ADSCs细胞浆内表达OCN及CAP。同时DFCCM诱导ADSCs后CAP,CP23,ALP及OCN mRNA表达水平显著高于对照组。结论:牙囊细胞条件培养液构建的微环境可使ADSCs向成牙骨质样细胞分化,为干细胞介导的牙骨质再生提供新的思路。

Objective： To explore the possibility of adipose derived stem cells （ADSCs） differentiation into ce- mentoblast--like cells. Methods： ADSCs were isolated from the inguinal region of 7--day--old rats and cultured in dental follicle cell conditioned media （DFCCM）. The proliferation of ADSCs was examined by MTT assay during a period of 8 d . After 7d induction, the protein and gene expression for cementum attachment protein （CAP） and mineralization--related markers （OCN, ALP） were assayed in ADSCs treated with DFCCM. Results： Compared with the control group, DFCCM treatment significantly inhibited the proliferation rate of ADSCs （P^0.05）. After 7d induction, expression of CAP and OCN was observed by immunofluorescence staining. Moreover, DFCCM treatment significantly induced the mRNA expression level of CAP, CP23, ALP and OCN compare to the control group （P%0.05）. Conclusion： Dental follicle cell conditioned media microenvieoment showed promotion effect on eementogenie differentiation of ADSCs. The results provide experimental evidence of tooth regeneration by ADSCs.