Generation of nephronal cells from human pluripotent stem cells via renal-vesicle formation

Human pluripotent stem cells (hPSC) provide a possible source for generation of kidney cells and organoids applicable in regenerative medicine, disease modeling and drug screening. By analyzing the effect of different factors on renal differentiation, we established an 8-day-protocol that steered hPSCs to the renal lineage by a step-wise process, segmented into mesoderm induction, intermediate mesoderm specification and metanephric induction outlining renal organogenesis. The differentiated cells contain populations of SIX2+/CITED1+ metanephric mesenchyme- (MM) and of HOXB7+/GRHL2+ ureteric bud (UB)-like cells at the end of 6 days. Transcriptome analysis corroborated that the in vitro generated precursor cell types at day 8 resemble their renal vesicle counterparts in vivo. The cells were further differentiated in 2-dimensional culture into podocyte- and diverse tubular epithelial-like cell types, showing their nephrogenic potential. In 3-dimensional culture, the progenitors gave rise to renal vesicles, and upon mouse embryonic kidney re-aggregation they form tubular structures.

人类多能干细胞（Human Pluripotent Stem Cells，hPSC）可为再生医学、疾病建模与药物筛选领域中应用的肾细胞及肾脏类器官的构建提供潜在来源。本研究通过分析不同因子对肾脏分化的调控作用，建立了一套为期8天的分步诱导方案，可将hPSC逐步定向诱导至肾谱系细胞；该诱导过程分为中胚层诱导、中间中胚层特化与后肾诱导三个阶段，完整勾勒肾脏发生的核心路径。诱导至第6天时，分化获得的细胞群体包含SIX2阳性/CITED1阳性的后肾间充质（metanephric mesenchyme，MM）样细胞，以及HOXB7阳性/GRHL2阳性的输尿管芽（ureteric bud，UB）样细胞。转录组分析证实，体外诱导第8天获得的前体细胞类型，与体内天然的肾囊泡前体细胞具有高度相似的分子特征。将该细胞群体在二维培养体系中进一步诱导，可分化为足细胞与多种肾小管上皮样细胞，证实其具备肾脏发生潜能。在三维培养体系中，这些前体细胞可形成肾囊泡；而将其与小鼠胚胎肾脏组织进行重聚集培养时，可进一步分化形成肾小管结构。