Despite a continuing increase in the number of patients suffering from chronic kidney disease,currently available treatments for these patients,such as dialysis and kidney transplantation,are imperfect.The kidney is a...Despite a continuing increase in the number of patients suffering from chronic kidney disease,currently available treatments for these patients,such as dialysis and kidney transplantation,are imperfect.The kidney is also a critical target organ vulnerable to the toxicity of various new drugs,and the lack of a reliable in vitro culture model imposes a severe limitation on drug discovery.Although the development of induced pluripotent stem cells(iPSCs)revolutionized strategies in biomedical fields,the complexity of the kidney imposed additional challenge to the application of this technology in kidney regeneration.Nonetheless,the recent advancement in our understanding on the developmental origin of kidney progenitor cells and the mechanisms of their reciprocal induction and self-organization has boosted research in kidney regeneration.Research since then has demonstrated that kidney organoids derived from iPSCs can serve as a useful model for drug discovery and toxicity screening,as well as for disease modeling,especially in combination with gene editing techniques.Moreover,attempts at kidney organoid implantation in animals have suggested their potential as an alternative source of kidney transplantation.In this review,we summarize recent progress on the generation of kidney organoids,as well as the obstacles that remain.展开更多
Human induced pluripotent stem cell(hiPSC)-derived kidney organoids have prospective applications ranging from basic disease modelling to personalised medicine.However,there remains a necessity to refine the bio-physi...Human induced pluripotent stem cell(hiPSC)-derived kidney organoids have prospective applications ranging from basic disease modelling to personalised medicine.However,there remains a necessity to refine the bio-physical and biochemical parameters that govern kidney organoid formation.Differentiation within fully-controllable and physiologically relevant 3D growth environments will be critical to improving organoid reproducibility and maturation.Here,we matured hiPSC-derived kidney organoids within fully synthetic self-assembling peptide hydrogels(SAPHs)of variable stiffness(storage modulus,G′).The resulting organoids con-tained complex structures comparable to those differentiated within the animal-derived matrix,Matrigel.Single-cell RNA sequencing(scRNA-seq)was then used to compare organoids matured within SAPHs to those grown within Matrigel or at the air-liquid interface.A total of 13,179 cells were analysed,revealing 14 distinct clusters.Organoid compositional analysis revealed a larger proportion of nephron cell types within Transwell-derived organoids,while SAPH-derived organoids were enriched for stromal-associated cell populations.Notably,dif-ferentiation within a higher G’SAPH generated podocytes with more mature gene expression profiles.Addi-tionally,maturation within a 3D microenvironment significantly reduced the derivation of off-target cell types,which are a known limitation of current kidney organoid protocols.This work demonstrates the utility of syn-thetic peptide-based hydrogels with a defined stiffness,as a minimally complex microenvironment for the selected differentiation of kidney organoids.展开更多
Kidney disease has become a global public health problem affecting over 750 million people worldwide and imposing a heavy economic burden on patients.The complex architecture of the human kidney makes it very difficul...Kidney disease has become a global public health problem affecting over 750 million people worldwide and imposing a heavy economic burden on patients.The complex architecture of the human kidney makes it very difficult to study the pathophysiology of renal diseases in vitro and to develop effective therapeutic options for patients.Even though cell lines and animal models have enriched our understanding,they fail to recapitulate key aspects of human kidney development and renal disease at cellular and functional levels.Organoids can be derived from either pluripotent stem cells or adult stem cells by strictly regulating key signalling pathways.Today,these self-differentiated organoids represent a promising technology to further understand the human kidney,one of the most complex organs,in an unprecedented way.The newly established protocols improved by organ-on-chip and coculture with immune cells will push kidney organoids towards the next generation.Herein,we focus on recent achievements in the application of kidney organoids in disease modelling,nephrotoxic testing,precision medicine,biobanking,and regenerative therapy,followed by discussions of novel strategies to improve their utility for biomedical research.The applications we discuss may help to provide new ideas in clinical fields.展开更多
Organoid models are used to study kidney physiology,such as the assessment of nephrotoxicity and underlying disease processes.Personalized human pluripotent stem cell-derived kidney organoids are ideal models for comp...Organoid models are used to study kidney physiology,such as the assessment of nephrotoxicity and underlying disease processes.Personalized human pluripotent stem cell-derived kidney organoids are ideal models for compound toxicity studies,but there is a need to accelerate basic and translational research in the field.Here,we developed an automated continuous imaging setup with the“read-on-ski”law of control to maximize temporal resolution with minimum culture plate vibration.High-accuracy performance was achieved:organoid screening and imaging were performed at a spatial resolution of 1.1µm for the entire multi-well plate under 3 min.We used the in-house developed multi-well spinning device and cisplatin-induced nephrotoxicity model to evaluate the toxicity in kidney organoids using this system.The acquired images were processed via machine learning-based classification and segmentation algorithms,and the toxicity in kidney organoids was determined with 95%accuracy.The results obtained by the automated“read-on-ski”imaging device,combined with label-free and non-invasive algorithms for detection,were verified using conventional biological procedures.Taking advantage of the close-to-in vivo-kidney organoid model,this new development opens the door for further application of scaled-up screening using organoids in basic research and drug discovery.展开更多
文摘Despite a continuing increase in the number of patients suffering from chronic kidney disease,currently available treatments for these patients,such as dialysis and kidney transplantation,are imperfect.The kidney is also a critical target organ vulnerable to the toxicity of various new drugs,and the lack of a reliable in vitro culture model imposes a severe limitation on drug discovery.Although the development of induced pluripotent stem cells(iPSCs)revolutionized strategies in biomedical fields,the complexity of the kidney imposed additional challenge to the application of this technology in kidney regeneration.Nonetheless,the recent advancement in our understanding on the developmental origin of kidney progenitor cells and the mechanisms of their reciprocal induction and self-organization has boosted research in kidney regeneration.Research since then has demonstrated that kidney organoids derived from iPSCs can serve as a useful model for drug discovery and toxicity screening,as well as for disease modeling,especially in combination with gene editing techniques.Moreover,attempts at kidney organoid implantation in animals have suggested their potential as an alternative source of kidney transplantation.In this review,we summarize recent progress on the generation of kidney organoids,as well as the obstacles that remain.
基金This publication has emanated from research conducted with the financial support of Science Foundation Ireland(SFI)co-funded under the European Regional Development Fund under Grant Number 13/RC/2073_P2+1 种基金The authors acknowledge support from Science Foundation Ireland(16/IA/4584)19/FFP/6833.J.K.W.would also like to acknowledge Royal Society of Chemistry grant(M19-6613).
文摘Human induced pluripotent stem cell(hiPSC)-derived kidney organoids have prospective applications ranging from basic disease modelling to personalised medicine.However,there remains a necessity to refine the bio-physical and biochemical parameters that govern kidney organoid formation.Differentiation within fully-controllable and physiologically relevant 3D growth environments will be critical to improving organoid reproducibility and maturation.Here,we matured hiPSC-derived kidney organoids within fully synthetic self-assembling peptide hydrogels(SAPHs)of variable stiffness(storage modulus,G′).The resulting organoids con-tained complex structures comparable to those differentiated within the animal-derived matrix,Matrigel.Single-cell RNA sequencing(scRNA-seq)was then used to compare organoids matured within SAPHs to those grown within Matrigel or at the air-liquid interface.A total of 13,179 cells were analysed,revealing 14 distinct clusters.Organoid compositional analysis revealed a larger proportion of nephron cell types within Transwell-derived organoids,while SAPH-derived organoids were enriched for stromal-associated cell populations.Notably,dif-ferentiation within a higher G’SAPH generated podocytes with more mature gene expression profiles.Addi-tionally,maturation within a 3D microenvironment significantly reduced the derivation of off-target cell types,which are a known limitation of current kidney organoid protocols.This work demonstrates the utility of syn-thetic peptide-based hydrogels with a defined stiffness,as a minimally complex microenvironment for the selected differentiation of kidney organoids.
基金This study was supported by grants from Undergraduate Inno-vation and Entrepreneurship Project(No.202321007)Hunan Innovative Provincial Construction Project(No.20195K2211)+7 种基金Changsha Science,Technology Plan Project(No.kq2001044)the National Natural Science Foundation of China(No.81970248)National Key Research and Development Program of China(No.2018YFA0108700,2017YFA0105602)NSFC Pro-jects of Intemational Cooperation and Exchanges(No.81720108004)National Natural Science Foundation of China(No.81974019)The Research Team Project of Natural Science Foundation of Guangdong Province of China(No.2017A030312007)The Key Program of Guangzhou Science Research Plan(No.201904020047)The Special Project of Dengfeng Program of Guangdong Provincial People's Hospital(No.DFJH201812,KJ012019119,KJ012019423).
文摘Kidney disease has become a global public health problem affecting over 750 million people worldwide and imposing a heavy economic burden on patients.The complex architecture of the human kidney makes it very difficult to study the pathophysiology of renal diseases in vitro and to develop effective therapeutic options for patients.Even though cell lines and animal models have enriched our understanding,they fail to recapitulate key aspects of human kidney development and renal disease at cellular and functional levels.Organoids can be derived from either pluripotent stem cells or adult stem cells by strictly regulating key signalling pathways.Today,these self-differentiated organoids represent a promising technology to further understand the human kidney,one of the most complex organs,in an unprecedented way.The newly established protocols improved by organ-on-chip and coculture with immune cells will push kidney organoids towards the next generation.Herein,we focus on recent achievements in the application of kidney organoids in disease modelling,nephrotoxic testing,precision medicine,biobanking,and regenerative therapy,followed by discussions of novel strategies to improve their utility for biomedical research.The applications we discuss may help to provide new ideas in clinical fields.
基金This research was funded by the Scientific Instrumentation Development Program of Chinese Academy of Sciences(No.ZDZBGCH2018005)the Key Research and Development Program of Bioland Laboratory(Guangzhou Regenerative Medicine and Health Guangdong Laboratory)(No.2019GZR1104060)the Research Instrument and Equipment Development Project of Chinese Academy of Sciences(No.ZDKYYQ20210006),China.
文摘Organoid models are used to study kidney physiology,such as the assessment of nephrotoxicity and underlying disease processes.Personalized human pluripotent stem cell-derived kidney organoids are ideal models for compound toxicity studies,but there is a need to accelerate basic and translational research in the field.Here,we developed an automated continuous imaging setup with the“read-on-ski”law of control to maximize temporal resolution with minimum culture plate vibration.High-accuracy performance was achieved:organoid screening and imaging were performed at a spatial resolution of 1.1µm for the entire multi-well plate under 3 min.We used the in-house developed multi-well spinning device and cisplatin-induced nephrotoxicity model to evaluate the toxicity in kidney organoids using this system.The acquired images were processed via machine learning-based classification and segmentation algorithms,and the toxicity in kidney organoids was determined with 95%accuracy.The results obtained by the automated“read-on-ski”imaging device,combined with label-free and non-invasive algorithms for detection,were verified using conventional biological procedures.Taking advantage of the close-to-in vivo-kidney organoid model,this new development opens the door for further application of scaled-up screening using organoids in basic research and drug discovery.
