TGF-β1介导人乳牙牙髓干细胞分化为血管平滑肌细胞的研究  

The study of functional smooth muscle cells differentiation from SHED induced by TGF-β1

作　　者：胡露露 艾琦[1,2] 胡晓燕 杨梓[1,2] 王银龙[1,2] 沈军[1,2] Hu Lulu1,2, Ai Qi1,2 , Hu Xiaoyan1,2 , et al(1 The Affiliated Stomatologic Hospital of Anhui Medical University, Key Laboratory of Oral Diseases Research of An,hui Province, Hefei 230032 ; 2Stomatologtc Hospital ＆ College, Anhui Medical University, Hefei 230032)

机构地区：[1]安徽医科大学口腔医学院,安徽省口腔疾病研究中心实验室,合肥230032 [2]安徽医科大学附属口腔医院正畸科,合肥230032

出　　处：《安徽医科大学学报》2018年第3期338-343,共6页Acta Universitatis Medicinalis Anhui

基　　金：安徽省自然科学基金资助计划(编号：KJ2011Z183;KJ2016A324)

摘　　要：目的血管化的主要方案是先生成血管组成细胞(血管内皮细胞和血管周细胞),然后将这些细胞构建成血管结构。由于受体自身内皮细胞和血管平滑肌细胞的稀缺性和异质性,限制了其在血管组织工程学中的广泛运用。因此,本研究对于人乳牙牙髓干细胞(SHED)是否能够诱导分化成为功能性血管平滑肌细胞(v SMCs)进行了相关方面的研究。方法利用转化生长因子(TGF)-β1作为刺激因子,诱导SHED分化为v SMCs。完成诱导后,通过RT-PCR、流式细胞术分析、Western blot和免疫荧光技术检测基因和特异性蛋白表达情况。此外,借助Matrigel血管生成功能实验来验证SHED来源的v SMCs是否能行使平滑肌细胞的功能。结果通过分析平滑肌细胞特定标志物(如α-SMA、SM22α、Calponin和SM-MHC)的表达,证实了TGF-β1可以诱导SHED分化为v SMCs,且流式细胞术证实分化的效率处于较高的水平,其标志物表达比例高达α-SMA 86.1%,SM22α93.9%,Calponin 56.8%,SM-MHC 88.2%。体外Matrigel血管生成功能实验表明,由SHED诱导分化的v SMCs在稳定由人脐静脉血管内皮细胞所形成的血管结构中发挥着与原代v SMCs相似的作用。结论在血管组织工程中,SHED可以成功地诱导为v SMCs,能够成为血管组织工程中一种有前景的种子细胞。Objective The main strategy of vascularization is to generate vascular cell （ endothelial cells and vascular smooth muscle cells）, and then recombine these cells into vascular structures. The scarcity and heterogenicity of primary endothelial ceils and vascular smooth muscle cells （vSMCs） hinder the advancement of vascular tissue engineering. Therefore, we investigated whether vSMCs could be generated from human exfoliated deciduous teeth （SHED）. Methods Transforming growth factor beta 1 （ TGF-β1 ） was utilized to induce SHED differentiation into vSMCs. The mRNA and protein expression were analyzed using RT-PCR, flow cytometry, Western blot and immu- nostaining. Furthermore, in vitro Matrigel angiogenesis assay was used to examine whether those SHED-derived vSMCs possess the similar function as primary SMCs. Results The results of analyzation of SMCs specific mark- ers＇ （SM22α, α-SMA, SM-MHC, and Calponin） expression confirmed that TGF-β1 could induce the differentia- tion of SMCs from SHED, and the efficiency of differentiation assessed by flow cytometry was relatively high （ α- SMA 86. 1%, SM22α 93.9%, Calponin 56. 8% and SM-MHC 88.2% ）. The results of in vitro Matrigel Angio- genesis Assay confirmed that SHED-derived SMCs possessed the similar function with primary SMCs in stabilizing the vascular structures generated by human umbilical vein endothelial cells. Conclusion SHED possesses the po- tential to generate functional SMCs, and could be a promising cell candidate in vascular tissue engineering.

关 键 词：组织工程学 成血管 乳牙牙髓干细胞 平滑肌细胞 

分 类 号：Q813.1[生物学—生物工程]