Pig and monkey are widely used models for exploration of human diseases and evaluation of drug efficiency and toxicity,but high cost limits their uses.Organoids have been shown to be promising models for drug test as ...Pig and monkey are widely used models for exploration of human diseases and evaluation of drug efficiency and toxicity,but high cost limits their uses.Organoids have been shown to be promising models for drug test as they reasonably preserve tissue structure and functions.However,colonic organoids of pig and monkey are not yet established.Here,we report a culture medium to support the growth of porcine and monkey colonic organoids.Wnt signaling and PGE2 are important for long-term expansion of the organoids,and their withdrawal results in lineage differentiation to mature cells.Furthermore,we observe that porcine colonic organoids are closer to human colonic organoids in terms of drug toxicity response.Successful establishment of porcine and monkey colonic organoids would facilitate the mechanistic investigation of the homeostatic regulation of the intestine of these animals and is useful for drug development and toxicity studies.展开更多
The recent advent of robust methods to grow human tissues as 3D organoids allows us to recapitulate the 3D architecture of tumors in an in vitro setting and offers a new orthogonal approach for drug discovery.However,...The recent advent of robust methods to grow human tissues as 3D organoids allows us to recapitulate the 3D architecture of tumors in an in vitro setting and offers a new orthogonal approach for drug discovery.However,organoid culturing with extracellular matrix to support 3D architecture has been challenging for high-throughput screening(HTS)-based drug discovery due to technical difficulties.Using genetically engineered human colon organoids as a model system,here we report our effort to miniaturize such 3D organoid culture with extracellular matrix support in high-density plates to enable HTS.We first established organoid culturing in a 384-well plate format and validated its application in a cell viability HTS assay by screening a 2036-compound library.We further miniaturized the 3D organoid culturing in a 1536-well ultra-HTS format and demonstrated its robust performance for large-scale primary compound screening.Our miniaturized organoid culturing method may be adapted to other types of organoids.By leveraging the power of 3D organoid culture in a high-density plate format,we provide a physiologically relevant screening platform to model tumors to accelerate organoid-based research and drug discovery.展开更多
基金This work was supported by grants from the National Key Research and Development Program of China(2017YFA0103601)the National Natural Science Foundation of China(31730056 and 31988101)to YGC.
文摘Pig and monkey are widely used models for exploration of human diseases and evaluation of drug efficiency and toxicity,but high cost limits their uses.Organoids have been shown to be promising models for drug test as they reasonably preserve tissue structure and functions.However,colonic organoids of pig and monkey are not yet established.Here,we report a culture medium to support the growth of porcine and monkey colonic organoids.Wnt signaling and PGE2 are important for long-term expansion of the organoids,and their withdrawal results in lineage differentiation to mature cells.Furthermore,we observe that porcine colonic organoids are closer to human colonic organoids in terms of drug toxicity response.Successful establishment of porcine and monkey colonic organoids would facilitate the mechanistic investigation of the homeostatic regulation of the intestine of these animals and is useful for drug development and toxicity studies.
基金This research was supported by the NCI Cancer TargetDiscovery and Development(CTD^2)Network(1U01CA217875 toH.F.and 1uo1CA217851 to C.J.K.)the RAS Synthetic LethalNetwork(RSLN+4 种基金1UO1CA199241 to C.J.K.)the Emory LungCancer SPORE(NIH P5OCA217691)the Winship Cancerlnstitute(NIH 5P30CA138292)the Emory WHSC 10x SingleCell Sequencing Seed Grant(X.M.and Y.D.)Emory WoodruffHealth Sciences Center Synergy Award,and the lmagine,lnnovate and Impact(3)Funds from the Emory School ofMedicine and through the Georgia CTSA NIH award(UL1-TRO02378).
文摘The recent advent of robust methods to grow human tissues as 3D organoids allows us to recapitulate the 3D architecture of tumors in an in vitro setting and offers a new orthogonal approach for drug discovery.However,organoid culturing with extracellular matrix to support 3D architecture has been challenging for high-throughput screening(HTS)-based drug discovery due to technical difficulties.Using genetically engineered human colon organoids as a model system,here we report our effort to miniaturize such 3D organoid culture with extracellular matrix support in high-density plates to enable HTS.We first established organoid culturing in a 384-well plate format and validated its application in a cell viability HTS assay by screening a 2036-compound library.We further miniaturized the 3D organoid culturing in a 1536-well ultra-HTS format and demonstrated its robust performance for large-scale primary compound screening.Our miniaturized organoid culturing method may be adapted to other types of organoids.By leveraging the power of 3D organoid culture in a high-density plate format,we provide a physiologically relevant screening platform to model tumors to accelerate organoid-based research and drug discovery.
