Application of human ES-derived vascular progenitor cells to elucidation of vascular development/differentiation and vascular regeneration medicine

我们从人ES细胞中鉴定并分离出了“血管祖细胞（VPC）”，其在体外可分化为内皮细胞（EC）和血管平滑肌细胞（VSMC）并组织血管结构。人ES细胞衍生的VPC（hESC-VPC）是TRA-1-Flk 1 +VECadherin-PDGFR+细胞。我们还发现，血管舒张肽，利钠肽和肾上腺髓质素（AM）可以激活cGMP和cAMP级联反应，促进血管再生，通过我们开发的几个基因工程小鼠，并通过腺病毒介导的基因转移技术。此外，我们证明了AM有效地诱导小鼠ES细胞来源的VPC的内皮分化。移植早期分化的EC和VSMC在体内诱导人血管的构建，并增强局部血流，以在后肢缺血模型或皮肤溃疡模型中发挥治疗作用。利用体外人血管发育系统，以人VPC为材料，研究了人未分化ES、VPC、EC和SMC在分化早期的基因表达谱。此外，我们还开发了一种新的血管壁生物材料，并将ES细胞来源的VPC接种于其上，在脉动流加载下，VPC在人工血管壁中分化为EC和VSMC。

From human ES cells we identified and isolated "vascular progenitor cells;VPC", which can be differentiated into both endothelial cells(EC) and vascular smooth muscle cells(VSMC) and organize blood vessel structure in vitro. Human ES cell-derived VPC(hESC-VPC) are TRA-1-Flk1+VEcadherin-PDGFR+ cells. We also discovered that vasodilating peptides, natriuretic peptides and adrenomedullin(AM) can activate cGMP and cAMP cascade, respectively and promote vascular regeneration, by several gene-engineered mice for these peptides, which we developed and by adenovirus-mediated gene transfer technique. Furthermore, we demonstrated that AM potently induces endothelial differentiation of mouse ES cell-derived VPC. Transplantation of EC and VSMC at early differentiation stage induced construction of human blood vessels in vivo and enhanced local blood flow to exert therapeutic effectiveness in hindlimb ischemia model or skin ulcer model. By in vitro human vascular development system, using human VPC, we elucidated the gene expression profiling of human undifferentiated ES,VPC and EC or SMC at early differentiation stage. Furthermore, we developed new biomaterial for blood vessel walls and seeded ES cell-derived VPC onto it. VPC in the artificial blood vessel walls were demonstrated to be differentiated into EC and VSMC by pulsatile flow loading.