血管平滑肌细胞(VSMCs)移植是促进血管新生和创伤愈合的一种有效的细胞治疗方法。然而VSMCs的来源不足严重局限了其在血管疾病治疗中的研究和应用。尽管目前人们对VSMCs分化机制的认识有所进展，但仍缺乏稳定高效的从多能干细胞中获得谱系特异性VSMCs的方法。我们在前期利用单细胞测序研究人脂肪间充质干细胞（ADSCs）的异质性时发现，ADSCs中存在VSMCs定向前体细胞的群体，且其表面存在与分化相关的细胞表面标记—S蛋白。同时，多个线索提示，将此群细胞纯化会显著提高ADSCs向VSMCs的分化效率。本课题将综合利用单细胞测序技术，生物信息学分析技术，细胞分选纯化技术，以及干细胞定向分化诱导技术，建立一种ADSCs—VSMC前体细胞亚群纯化—VSMC定向分化的标准化诱导流程，并阐述S蛋白在VSMC定向分化中的分子调控机制；为血管疾病的细胞治疗提供大量有功能的成熟VSMC细胞。

Transplantation of vascular smooth muscle cells (VSMCs) is a promising cellular therapy to promote angiogenesis and wound healing. However, the insufficiency of VSMCs greatly limits their therapeutic application in vascular disease. Despite progress in understanding the mechanisms of VSMC differentiation, there remains a shortage of robust methods for generating lineage-specific VSMCs from pluripotent and adult stem/progenitor cells in serum-free conditions. Recently, using single cell sequencing to study the heterogeneity of ADSCs, we found that there existed a group of VSMCs oriented precursor cells in adipose derived mesenchymal stem cells (ADSCs). Furthermore, several clues suggest that the purification of these cells may significantly increase the differentiation efficiency of ADSCs into lineage-specific VSMCs. In this study, we devote to establish a standardized induction process for ADSCs - VSMC precursor cell subsets purification - VSMC directional differentiation, and expound the molecular regulatory mechanism underlying S protein-related directional differentiation of VSMC by the integrated application of single cell sequencing, bioinformatics analysis, cell separation and purification, and directional differentiation of stem cells. This work will not only enable us to provide sufficient functional VSMC–like cells for cell therapy of vascular disease, but also shed lights in understanding the molecular mechanisms of VSMC precursor cell directional differentiation.