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Chemically defined and scalable culture system for intestinal stem cells derived from human intestinal organoids (RNA-Seq)

ABSTRACT: we established a human intestinal stem cell (ISC) culture method expanded under feeder-free and fully defined conditions through selective enrichment of ISC populations (ISC3D-hIO) within hIO derived from human pluripotent stem cells.

ORGANISM(S): Homo sapiens

PROVIDER: GSE219016 | GEO | 2023/11/20

REPOSITORIES: GEO

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Chemically defined and scalable culture system for intestinal stem cells derived from human intestinal organoids (scRNA-Seq)

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2023-11-20 | GSE219017 | GEO

Xenogeneic-free culture of human intestinal stem cells on functional polymer-coated substrates for scalable, clinical-grade stem cell therapy

Project description:Inevitable need for basement membrane extract (BME) with undefined constituents, such as Matrigel, for intestinal stem cell (ISC) culture in traditional system poses a significant barrier that must be overcome for the development of clinical-grade, scalable, ready-to-use ISCs. Here, we propose a functional polymer-based xenogeneic-free dish for the culture of intestinal stem cells (XF-DISC), ensuring substantially prolonged maintenance of ISCs derived from 3-dimensional human intestinal organoid (ISCs3D-hIO). XF-DISC enables remarkable expandability, exhibiting a 24-fold amplification in cell number within 30 days, with long-term maintenance of ISCs3D-hIO through consecutive passaging more than 30 times (> 210 days), and is fully compatible with a cell banking system. Notably, the ISCs3D-hIO cultured on XF-DISC are successfully transplanted into the intestinal injury and inflammation models, leading to full regeneration of damaged intestinal epithelium. XF-DISC, an extraordinary reliable and scalable xenogeneic-free ISC3D-hIO culture method exhibits significant promise in the development of regenerative ISC therapy for human intestinal diseases.

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Chemically-defined and scalable culture system for intestinal stem cells derived from human intestinal organoids.

Project description:Three-dimensional human intestinal organoids (hIO) are widely used as a platform for biological and biomedical research. However, reproducibility and challenges for large-scale expansion limit their applicability. Here, we establish a human intestinal stem cell (ISC) culture method expanded under feeder-free and fully defined conditions through selective enrichment of ISC populations (ISC3D-hIO) within hIO derived from human pluripotent stem cells. The intrinsic self-organisation property of ISC3D-hIO, combined with air-liquid interface culture in a minimally defined medium, forces ISC3D-hIO to differentiate into the intestinal epithelium with cellular diversity, villus-like structure, and barrier integrity. Notably, ISC3D-hIO is an ideal cell source for gene editing to study ISC biology and transplantation for intestinal diseases. We demonstrate the intestinal epithelium differentiated from ISC3D-hIO as a model system to study severe acute respiratory syndrome coronavirus 2 viral infection. ISC3D-hIO culture technology provides a biological tool for use in regenerative medicine and disease modelling.

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Chemically defined and scalable culture system for intestinal stem cells derived from human intestinal organoids

Project description:This SuperSeries is composed of the SubSeries listed below.

2023-11-20 | GSE219018 | GEO

Chemically defined and scalable culture system for intestinal stem cells derived from human intestinal organoids

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Chemically defined and scalable culture system for intestinal stem cells derived from human intestinal organoids (scRNA-Seq)

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Project description:Tissue organoids are a promising technology that may accelerate development of the societal and NIH mandate for precision medicine. Here we describe a robust and simple method for generating cerebral organoids (cOrgs) from human pluripotent stem cells by using a chemically defined hydrogel material and chemically defined culture medium. By using no additional neural induction components, cOrgs appeared on the hydrogel surface within 10-14 days, and under static culture conditions, they attained sizes up to 3 mm in greatest dimension by day 28. Histologically, the organoids showed neural rosette and neural tube-like structures and evidence of early corticogenesis. Immunostaining and quantitative reverse-transcription polymerase chain reaction demonstrated protein and gene expression representative of forebrain, midbrain, and hindbrain development. Physiologic studies showed responses to glutamate and depolarization in many cells, consistent with neural behavior. The method of cerebral organoid generation described here facilitates access to this technology, enables scalable applications, and provides a potential pathway to translational applications where defined components are desirable.Tissue organoids are a promising technology with many potential applications, such as pharmaceutical screens and development of in vitro disease models, particularly for human polygenic conditions where animal models are insufficient. This work describes a robust and simple method for generating cerebral organoids from human induced pluripotent stem cells by using a chemically defined hydrogel material and chemically defined culture medium. This method, by virtue of its simplicity and use of defined materials, greatly facilitates access to cerebral organoid technology, enables scalable applications, and provides a potential pathway to translational applications where defined components are desirable.

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