Parallel generation of easily selectable multiple nephronal cell types from human pluripotent stem cells
Parallel generation of easily selectable multiple nephronal cell types from human pluripotent stem cells
复制标题
DOI：
10.1007/s00018-018-2929-2
复制
发表时间：
2019-01-01
影响因子：
8
通讯作者：
Reinke, Petra
中科院分区：
文献类型：
--
作者：
Hariharan, Krithika;Stachelscheid, Harald;Reinke, Petra
人多能干细胞 (hPSC) 为生成适用于再生医学、疾病建模和药物筛选的特定肾细胞和肾类器官提供了来源。这些应用需要通过可重复、可扩展且有效的方法提供 hPSC 衍生的肾细胞。我们通过应用激活素 A、BMP4 和视黄酸，然后应用 GDNF 建立了化学定义的方案，该方案将 hPSC 引导至肾谱系，并在 6 天时产生 SIX2(+)/CITED1(+) 后肾间充质 (MM) 和 HOXB7(+)/GRHL2(+) 输尿管芽 (UB) 样细胞群。转录组分析证实，第 8 天时的 PSC 衍生细胞类型类似于体内的肾小泡和输尿管上皮细胞，6 天后形成肾小管和肾小球细胞。我们证明，从 hPSC 开始，我们的体外方案在肾囊泡阶段生成了肾源祖细胞库，14 天后可以进一步定向为特殊的肾单位细胞类型，包括系膜细胞、近端肾小管细胞、远端肾小管细胞、集合管上皮细胞和足细胞前体细胞。这种简单而快速的方法可以在 2D 培养中从常见的前体池中产生肾细胞，为扩大生产定制的肾细胞类型提供了基础，这些细胞适用于药物测试或基于细胞的再生疗法。
Human pluripotent stem cells (hPSCs) provide a source for the generation of defined kidney cells and renal organoids applicable in regenerative medicine, disease modeling, and drug screening. These applications require the provision of hPSC-derived renal cells by reproducible, scalable, and efficient methods. We established a chemically defined protocol by application of Activin A, BMP4, and Retinoic acid followed by GDNF, which steered hPSCs to the renal lineage and resulted in populations of SIX2(+)/CITED1(+) metanephric mesenchyme- (MM) and of HOXB7(+)/GRHL2(+) ureteric bud (UB)-like cells already by 6days. Transcriptome analysis corroborated that the PSC-derived cell types at day 8 resemble their renal vesicle and ureteric epithelial counterpart in vivo, forming tubular and glomerular renal cells 6 days later. We demonstrate that starting from hPSCs, our in vitro protocol generates a pool of nephrogenic progenitors at the renal vesicle stage, which can be further directed into specialized nephronal cell types including mesangial-, proximal tubular-, distal tubular, collecting duct epithelial cells, and podocyte precursors after 14days. This simple and rapid method to produce renal cells from a common precursor pool in 2D culture provides the basis for scaled-up production of tailored renal cell types, which are applicable for drug testing or cell-based regenerative therapies.