Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, Singapore, 138669, Singapore.
Adv Healthc Mater. 2016 Aug;5(16):2080-91. doi: 10.1002/adhm.201600120. Epub 2016 Jun 13.
Recent advances in developmental biology and stem cell technology have led to the engineering of functional organs in a dish. However, the limited size of these organoids and absence of a large circulatory system poses limits to its clinical translation. To overcome these issues, decellularized whole kidney scaffolds with native microstructure and extracellular matrix (ECM) are employed for kidney bioengineering, using human-induced pluripotent-stem-cell-derived renal progenitor cells and endothelial cells. To demonstrate ECM-guided cellular assembly, the present work is focused on generating the functional unit of the kidney, the glomerulus. In the repopulated organ, the presence of endothelial cells broadly upregulates the expression level of genes related to renal development. When the cellularized native scaffolds are implanted in SCID mice, glomeruli assembly can be achieved by co-culture of the renal progenitors and endothelial cells. These individual glomerular units are shown to be functional in the context of the whole organ using a simulated bio-reactor set-up with urea and creatinine excretion and albumin reabsorption. Our results indicate that the repopulation of decellularized native kidney using clinically relevant, expandable patient-specific renal progenitors and endothelial cells may be a viable approach for the generation of a functional whole kidney.
近年来，发育生物学和干细胞技术的进步使得在培养皿中构建功能性器官成为可能。然而，这些类器官的大小有限，且缺乏大型循环系统，这限制了它们在临床上的转化应用。为了克服这些问题，人们利用去细胞化的全肾支架，结合天然的微观结构和细胞外基质（ECM），使用人诱导多能干细胞衍生的肾祖细胞和内皮细胞进行肾脏生物工程。为了展示 ECM 引导的细胞组装，本工作专注于生成肾脏的功能单位——肾小球。在再定植的器官中，内皮细胞的广泛存在会大大上调与肾脏发育相关的基因的表达水平。当将细胞化的天然支架植入 SCID 小鼠体内时，通过共培养肾祖细胞和内皮细胞，可实现肾小球的组装。在使用模拟生物反应器的设置，进行尿素和肌酐排泄以及白蛋白重吸收的情况下，这些单个的肾小球单位在整个器官中表现出功能。我们的结果表明，使用临床上相关的、可扩增的患者特异性肾祖细胞和内皮细胞对去细胞化的天然肾脏进行再定植，可能是生成功能性全肾的一种可行方法。
