摘要
肾是一种重要的人体器官,具有多种生理功能。然而,全球范围内约有10%的人口患有肾疾病。因此,建立一种接近人体肾的结构与功能的模型进行肾疾病的研究是十分必要的。多能干细胞体外定向诱导分化技术的兴起,为再生医学和精准医学领域注入了新的动力。本研究通过在体外条件下模拟体内肾发育的过程,将人多能干细胞包括胚胎干细胞和诱导多能干细胞,通过体外定向诱导分化形成肾的祖细胞,进而建立肾的结构与功能单位:肾元。该研究通过激活WNT信号通路,同时抑制TGF-β信号通路,将人多能干细胞从多能态定向诱导至原条阶段。之后通过细胞自分化的能力使其发育至中间中胚层,再通过激活FGF信号通路,将其分化至肾祖细胞阶段。流式细胞检测结果显示,肾祖细胞占总细胞数的51.5%~61.9%。通过免疫荧光检测发现:分化得到的结构中包含肾小球足细胞、近端小管、远端小管等肾组织结构。该研究建立的肾体外分化方法,具有稳定性好、分化效率高、重复性好的特点。为研究人类肾的早期发育机制,肾疾病模型构建,以及药物筛选提供了一种新的方法。
Kidney is an essential organ in human body with multiple physiological functions.However,there is 10%population worldwide with renal disease.It is urgent to generate a model which is more similar with kidney at structural and functional level to study renal disease.The rise of in vitro differentiation technology from pluripotent stem cells gives regeneration medicine and precise medicine new energy.This study mimics kidney development in vitro by inducing human pluripotent stem cells including embryonic stem cells(ESCs)and induced pluripotent stem cells(iPSCs)into kidney progenitor cells,and further forming nephrons,which is the structure and function unit in kidney.Human pluripotent stem cells were differentiated into primitive streak through activating WNT pathway while inhibiting TGF-βsignaling.Afterward,the primitive streak spontaneously differentiated into intermediate mesoderm.Then,we induced intermediate mesoderm cells into kidney progenitor cells through FGF pathway.The FACS analysis data indicated kidney progenitor cells were up to 51.5%-61.9%in total cell population.Immuno-staining results showed these structures contained podocytes of glomerulus,proximal tubule,and distal tubule.This kidney differentiation protocol is stable,high-efficient,and well repeatable.This research provides a novel platform for early human kidney development study,kidney disease modeling,and drug screening.
作者
周冰蕊
魏云亮
梁婷
何生
冯志伟
靳宁
刘志贞
赵虹
侯淑琳
于保锋
解军
ZHOU Bing-Rui;WEI Yun-Liang;LIANG Ting;HE Sheng;FENG Zhi-Wei;JIN Ning;LIU Zhi-Zhen;ZHAO Hong;HOU Shu-Lin;YU Bao-Feng;XIE Jun(Shanxi Key Laboratory of Birth Defect and Cell Regeneration,Department of Biochemistry and Molecular Biology,Shanxi Medical University;Department of Radiology,First Hospital of Shanxi Medical University,Taiyuan 030001,China)
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2022年第8期1098-1105,共8页
Chinese Journal of Biochemistry and Molecular Biology
基金
国家自然科学基金(No.82100821)
山西省重点研发计划(No.201903D421023)
中央引导地方科技发展资金项目(No.YDZJSX2021B008)
山西省基础研究计划(No.20210302124406)
山西医科大学博士启动基金项目(No.SD2012,No.XD2019)资助
关键词
人多能干细胞
体外诱导分化
肾的分化
肾元
human pluripotent stem cell
in vitro induced differentiation
kidney differentiation
nephron
