Background:Intestinal organoids are promising tools in the context of animal experiment reduction but a thorough characterization of the impact of the origin of intestinal stem cells(ISC)on organoid phenotype is neede...Background:Intestinal organoids are promising tools in the context of animal experiment reduction but a thorough characterization of the impact of the origin of intestinal stem cells(ISC)on organoid phenotype is needed to routinely use this cellular model.Our objective was to evaluate the effect of ISC donor age on the growth,morphology and cellular composition of intestinal organoids derived from pig.Methods:Organoids were derived from jejunal and colonic ISC obtained from 1-,7-,28-,36-and 180-day-old pigs and passaged three times.Results:We first confirmed by qPCR that the expression of 18%of the>80 studied genes related to various intestinal functions differed between jejunal and colonic organoids after two passages(p<0.05).Growth and morphology of organoids depended on intestinal location(greater number and larger organoids derived from colonic than jejunal ISC,p<0.05)but also pig age.Indeed,when ISC were derived from young piglets,the ratio of organoids to spheroids was greater(p<0.05),spheroids were larger during the primary culture but smaller after two passages(p<0.05)and organoids were smaller after one passage(p>0.05)compared to ISC from older pigs.Finally,no difference in cellular composition,evaluated by immunostaining of markers of the major intestinal cell types(absorptive,enteroendocrine and goblet cells)was observed between organoids originating from 7-or 180-day-old pigs,but differences between intestinal site origins were noticed.Conclusion:In conclusion,while the age of the tissue donor affected organoid growth and morphology,it did not influence the phenotype.展开更多
Intestinal epithelial stem cells (IESCs) are one of the most rapidly self‐renewing and proliferating adult stem cells. The IESCs reside at the bottom of intestinal and colonic crypts, giving rise to all intestinal ep...Intestinal epithelial stem cells (IESCs) are one of the most rapidly self‐renewing and proliferating adult stem cells. The IESCs reside at the bottom of intestinal and colonic crypts, giving rise to all intestinal epithelial lineages and maintaining intestinal epithelial replenishment. The technique of three‐dimensional culture based upon intestinal stem cell biology has been recently developed to study gastrointestinal development and disease pathogenesis. Here, we summarize the techniques used to isolate Lgr5‐ positive IESCs to form the enteroids from intestine or colonoids from colon, and present the means to examine these organoid functions. This study will provide a simple and practical way for producing intestinal tissues in the laboratory.展开更多
BACKGROUND： Organoid is an in vitro three-dimensional organ-bud that shows realistic microanatomy and physiological relevance. The progress in generating organoids that faithfully recapitulate human in vivo tissue co...BACKGROUND： Organoid is an in vitro three-dimensional organ-bud that shows realistic microanatomy and physiological relevance. The progress in generating organoids that faithfully recapitulate human in vivo tissue composition has extended organoid applications from being just a basic research tool to a translational platform with a wide range of uses. Study of host- microbial interactions relies on model systems to mimic the in vivo infection. Researchers have developed various experimental models in vitro and in vivo to examine the dynamic host-microbial interactions. For some infectious pathogens, model systems are lacking whereas some of the used systems are far from optimal. OBJECTIVE： In the present work, we will review the brief history and recent findings using organoids for studying host- microbial interactions. METHODS： A systematic literature search was performed using the PubMed search engine. We also shared our data and research contribution to the field. RESULTS： we summarize the brief history of 3D organoids. We discuss the feasibility of using organoids in studying host- microbial interactions, focusing on the development of intestinal organoids and gastric organoids. We highlight the advantage and challenges of the new experimental models. Further, we discuss the future direction in using organoids in studying host- microbial interactions and its potential application in biomedical studies. CONCLUSION： In combination with genetic, transcriptome and proteomic profiling, both murine- and human-derived organoids have revealed crucial aspects of development, homeostasis and diseases. Specifically, human organoids from susceptible host will be used to test their responses to pathogens, probiotics, and drugs. Organoid system is an exciting tool for studying infectious disease, microbiome, and therapy.展开更多
文摘Background:Intestinal organoids are promising tools in the context of animal experiment reduction but a thorough characterization of the impact of the origin of intestinal stem cells(ISC)on organoid phenotype is needed to routinely use this cellular model.Our objective was to evaluate the effect of ISC donor age on the growth,morphology and cellular composition of intestinal organoids derived from pig.Methods:Organoids were derived from jejunal and colonic ISC obtained from 1-,7-,28-,36-and 180-day-old pigs and passaged three times.Results:We first confirmed by qPCR that the expression of 18%of the>80 studied genes related to various intestinal functions differed between jejunal and colonic organoids after two passages(p<0.05).Growth and morphology of organoids depended on intestinal location(greater number and larger organoids derived from colonic than jejunal ISC,p<0.05)but also pig age.Indeed,when ISC were derived from young piglets,the ratio of organoids to spheroids was greater(p<0.05),spheroids were larger during the primary culture but smaller after two passages(p<0.05)and organoids were smaller after one passage(p>0.05)compared to ISC from older pigs.Finally,no difference in cellular composition,evaluated by immunostaining of markers of the major intestinal cell types(absorptive,enteroendocrine and goblet cells)was observed between organoids originating from 7-or 180-day-old pigs,but differences between intestinal site origins were noticed.Conclusion:In conclusion,while the age of the tissue donor affected organoid growth and morphology,it did not influence the phenotype.
基金The Non-profit Central Institute Fund of Chinese Academy of Medical Sciences,Grant/Award Number:2016RC310011The National Key Research and Development Program of China,Grant/Award Number:2017YFA0105200Peking Medical College(PUMC)Scholar-Endowed Professorship
文摘Intestinal epithelial stem cells (IESCs) are one of the most rapidly self‐renewing and proliferating adult stem cells. The IESCs reside at the bottom of intestinal and colonic crypts, giving rise to all intestinal epithelial lineages and maintaining intestinal epithelial replenishment. The technique of three‐dimensional culture based upon intestinal stem cell biology has been recently developed to study gastrointestinal development and disease pathogenesis. Here, we summarize the techniques used to isolate Lgr5‐ positive IESCs to form the enteroids from intestine or colonoids from colon, and present the means to examine these organoid functions. This study will provide a simple and practical way for producing intestinal tissues in the laboratory.
文摘BACKGROUND： Organoid is an in vitro three-dimensional organ-bud that shows realistic microanatomy and physiological relevance. The progress in generating organoids that faithfully recapitulate human in vivo tissue composition has extended organoid applications from being just a basic research tool to a translational platform with a wide range of uses. Study of host- microbial interactions relies on model systems to mimic the in vivo infection. Researchers have developed various experimental models in vitro and in vivo to examine the dynamic host-microbial interactions. For some infectious pathogens, model systems are lacking whereas some of the used systems are far from optimal. OBJECTIVE： In the present work, we will review the brief history and recent findings using organoids for studying host- microbial interactions. METHODS： A systematic literature search was performed using the PubMed search engine. We also shared our data and research contribution to the field. RESULTS： we summarize the brief history of 3D organoids. We discuss the feasibility of using organoids in studying host- microbial interactions, focusing on the development of intestinal organoids and gastric organoids. We highlight the advantage and challenges of the new experimental models. Further, we discuss the future direction in using organoids in studying host- microbial interactions and its potential application in biomedical studies. CONCLUSION： In combination with genetic, transcriptome and proteomic profiling, both murine- and human-derived organoids have revealed crucial aspects of development, homeostasis and diseases. Specifically, human organoids from susceptible host will be used to test their responses to pathogens, probiotics, and drugs. Organoid system is an exciting tool for studying infectious disease, microbiome, and therapy.
