Perivascular Progenitor Cells Derived from Human Embryonic Stem Cells Exhibit Functional Characteristics of Pericytes, and Improve the Retinal Vasculature in a Rodent Model of Diabetic Retinopathy

Diabetic retinopathy (DR) is the leading cause of blindness in working-age people. Pericyte loss is one of the pathologic cellular events in DR, which weakens the retinal microvessels. Damages to the microvascular networks are irreversible and permanent, thus further progression of DR is inevitable. In this study, we hypothesize that multipotent perivascular progenitor cells derived from human ESCs (hESC-PVPCs) improve the damaged retinal vasculature in the streptozotocin (STZ)-induced diabetic rodent models. We describe a highly efficient and feasible protocol to derive such cells using a natural selection method without cell sorting processes. As a cellular model of pericytes, hESC-PVPCs exhibited marker expressions such as CD140B, CD146, NG2, and functional characteristics of pericytes. Following a single intravitreal injection into diabetic Brown Norway (BN) rats, we demonstrate that the cells localized alongside typical perivascular regions of the retinal vasculature, and stabilized blood-retinal barrier (BRB) breakdown. Findings in this study highlight a therapeutic potential of hESC-PVPCs in DR by mimicking the role of pericytes in vascular stabilization. The molecular profiling analysis of ontological selected gene sets demonstrated that the transcripts associated with stemness markers, neural crest, neuro-ectoderm, and endoderm gene expressions.

糖尿病视网膜病变（Diabetic retinopathy, DR）是工作年龄人群首要的致盲病因。周细胞（pericyte）丢失是DR的病理性细胞事件之一，可削弱视网膜微血管的结构与功能。微血管网络的损伤具有不可逆性与永久性，因此DR的病情将不可避免地进一步进展。本研究提出假说：人类胚胎干细胞来源的多能血管周祖细胞（multipotent perivascular progenitor cells, hESC-PVPCs）可改善链脲佐菌素（streptozotocin, STZ）诱导的糖尿病啮齿动物模型中受损的视网膜血管系统。我们开发了一种基于自然筛选、无需细胞分选流程的高效可行方案，以获取此类细胞。作为周细胞的体外模型，hESC-PVPCs可表达CD140B、CD146、NG2等周细胞标志物，并具备周细胞的功能特性。向糖尿病布朗挪威（Brown Norway, BN）大鼠实施单次玻璃体内注射后，我们证实该细胞可定位于视网膜血管的典型血管周区域，并缓解血视网膜屏障（blood-retinal barrier, BRB）的破坏。本研究结果表明，hESC-PVPCs通过模拟周细胞维持血管稳定的生理作用，在DR治疗中具有潜在的应用价值。对本体筛选得到的基因集进行分子谱分析（molecular profiling analysis）显示，该细胞的转录本与干性标志物（stemness markers）、神经嵴（neural crest）、神经外胚层（neuro-ectoderm）及内胚层（endoderm）的基因表达特征相关。