Human Intestinal Organoids Group Standard 公告 2024-12-16 08:48:51 Written By Kym Kilbourne Title: Human Intestinal Organoids Group Standard Abstract The group standard "Human Intestinal Organoids" (T/CSCB0005—2022) is issued by the Chinese Society for Cell Biology, stipulating the ethical requirements, technical requirements, and testing methods for human intestinal organoids. This standard applies to the in vitro preparation and testing of organoids derived from human intestinal epithelial tissue, aiming to regulate the research and application of organoids, ensuring their scientific and standardized practice. Standard Content Scope This document specifies the definition, ethical, and technical requirements, as well as testing methods for human intestinal organoids, providing standardized guidance for their preparation and testing. Normative References Includes WS213 diagnostic standards for Hepatitis B, WS293 diagnostic standards for AIDS and HIV infection, WS299 diagnostic standards for Hepatitis C, and the People's Republic of China Pharmacopoeia (2020 edition), ensuring the safety and effectiveness of organoid preparation and testing. Terms and Definitions Defines key terms such as organoids, human intestinal organoids, passage, cryopreservation, revival, intestinal stem cells, intestinal stem cell differentiation, transient amplifying cells, absorptive enterocytes, goblet cells, Paneth cells, and enteroendocrine cells. Abbreviations Includes abbreviations such as Ct, DAPI, DMSO, HBV, HCV, HIV, PBS, PCR, RNA, STR, etc., facilitating the rapid identification and use of professional terminology. Ethical Requirements Requires the signing of an informed consent form with the organoid tissue donor, protection of donor personal information, and ensures that the research protocol is reviewed by an ethics committee. Technical Requirements 6.1 Morphology Organoids should appear as cystic or bud-like structures under an optical microscope, with lumens and closely packed columnar epithelial cells. 6.2 Karyotype The normal karyotype should be 46, XY or 46, XX. 6.3 Marker Genes Expression of marker genes such as LGR5, MUC2, ALPI, CHGA, MKI67 should be detected. 6.4 Cellular Composition Organoids should contain various types of intestinal epithelial cells, including intestinal stem cells, transient amplifying cells, absorptive enterocytes, goblet cells, Paneth cells, and enteroendocrine cells. 6.5 Functional Markers Functional markers such as alkaline phosphatase and lysozyme should be detected. 6.6 In Vitro Culture and Growth Organoids should be able to passage at least 5 generations in vitro and maintain the ability for self-renewal and intestinal stem cell differentiation. 6.7 Viability The viability of newly revived organoids should not be lower than 50%. 6.8 Microorganisms Organoids should test negative for fungi, bacteria, mycoplasmas, HBV, HCV, HIV, and exogenous viral factors. 6.9 STR Organoids cultured in vitro should be consistent with the donor's STR. Testing Methods Detailed descriptions of the testing methods for morphology, karyotype, marker genes, cellular composition, functional markers, quantity, viability, microorganisms, and STR are provided. Appendix Includes specific experimental steps and methods for the detection of cell type marker genes, the proportion of various cell compositions, alkaline phosphatase detection, mucin detection, organoid viability testing, and organoid STR identification. Conclusion The issuance of the "Human Intestinal Organoids" group standard provides a standardized framework for the research and application of human intestinal organoids, helping to improve the quality and reliability of organoid preparation and testing, and promoting the advancement of intestinal disease research and treatment. Kym Kilbourne
Human Intestinal Organoids Group Standard 公告 2024-12-16 08:48:51 Written By Kym Kilbourne Title: Human Intestinal Organoids Group Standard Abstract The group standard "Human Intestinal Organoids" (T/CSCB0005—2022) is issued by the Chinese Society for Cell Biology, stipulating the ethical requirements, technical requirements, and testing methods for human intestinal organoids. This standard applies to the in vitro preparation and testing of organoids derived from human intestinal epithelial tissue, aiming to regulate the research and application of organoids, ensuring their scientific and standardized practice. Standard Content Scope This document specifies the definition, ethical, and technical requirements, as well as testing methods for human intestinal organoids, providing standardized guidance for their preparation and testing. Normative References Includes WS213 diagnostic standards for Hepatitis B, WS293 diagnostic standards for AIDS and HIV infection, WS299 diagnostic standards for Hepatitis C, and the People's Republic of China Pharmacopoeia (2020 edition), ensuring the safety and effectiveness of organoid preparation and testing. Terms and Definitions Defines key terms such as organoids, human intestinal organoids, passage, cryopreservation, revival, intestinal stem cells, intestinal stem cell differentiation, transient amplifying cells, absorptive enterocytes, goblet cells, Paneth cells, and enteroendocrine cells. Abbreviations Includes abbreviations such as Ct, DAPI, DMSO, HBV, HCV, HIV, PBS, PCR, RNA, STR, etc., facilitating the rapid identification and use of professional terminology. Ethical Requirements Requires the signing of an informed consent form with the organoid tissue donor, protection of donor personal information, and ensures that the research protocol is reviewed by an ethics committee. Technical Requirements 6.1 Morphology Organoids should appear as cystic or bud-like structures under an optical microscope, with lumens and closely packed columnar epithelial cells. 6.2 Karyotype The normal karyotype should be 46, XY or 46, XX. 6.3 Marker Genes Expression of marker genes such as LGR5, MUC2, ALPI, CHGA, MKI67 should be detected. 6.4 Cellular Composition Organoids should contain various types of intestinal epithelial cells, including intestinal stem cells, transient amplifying cells, absorptive enterocytes, goblet cells, Paneth cells, and enteroendocrine cells. 6.5 Functional Markers Functional markers such as alkaline phosphatase and lysozyme should be detected. 6.6 In Vitro Culture and Growth Organoids should be able to passage at least 5 generations in vitro and maintain the ability for self-renewal and intestinal stem cell differentiation. 6.7 Viability The viability of newly revived organoids should not be lower than 50%. 6.8 Microorganisms Organoids should test negative for fungi, bacteria, mycoplasmas, HBV, HCV, HIV, and exogenous viral factors. 6.9 STR Organoids cultured in vitro should be consistent with the donor's STR. Testing Methods Detailed descriptions of the testing methods for morphology, karyotype, marker genes, cellular composition, functional markers, quantity, viability, microorganisms, and STR are provided. Appendix Includes specific experimental steps and methods for the detection of cell type marker genes, the proportion of various cell compositions, alkaline phosphatase detection, mucin detection, organoid viability testing, and organoid STR identification. Conclusion The issuance of the "Human Intestinal Organoids" group standard provides a standardized framework for the research and application of human intestinal organoids, helping to improve the quality and reliability of organoid preparation and testing, and promoting the advancement of intestinal disease research and treatment. Kym Kilbourne
