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Transcriptome analysis of interleukin-4 induced intestinal organoids from Japanese monkey

ABSTRACT: Several chemosensory cells exist throughout gastrointestinal tracts to sense nutrition and contaminants such as bacteria or parasites. Although rodent tissues are extensively used as a human gut model, recent studies show that the chemical sensing system in rodents differs from that in humans. Thus, experiments using rodents would not be the first choice. Nonhuman primates in biomedical research are valuable animal models to advance our understanding of biological responses in humans since availability of human tissues are limited. It has been reported that the 3D organoid culture produces functional gastrointestinal epithelial cells in vitro and can be generated from animal tissues. We have generated intestinal chemosensory cells from Japanese monkey using an organoid culture system. We could maintain monkey intestinal organoids in the proliferation medium for over one year. In this study, we aim to see changes in gene expression when monkey intestinal organoids were stimulated by interleukin (IL)-4. Previous studies revealed that these intestinal organoids express cytokine receptors such as IL-4R and response to IL-4 supplementation. We stimulated organoids with IL-4 for 72 hrs followed by total RNA purification and RNAs were subjected to transcriptome analysis

ORGANISM(S): Macaca fuscata

PROVIDER: GSE178817 | GEO | 2021/07/01

REPOSITORIES: GEO

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Project description:Background & Aims Patient-derived gastrointestinal organoids are powerful tools for studying human physiology and disease. However, generating organoids requires prompt isolation and culture of fresh tissue, which is labor and time intensive, placing restraints on basic and clinical research. For example organoid biobanking efforts, or the ability to obtain specimens from distant locations that lack the expertise to culture organoids, is limited by current approaches. We sought to develop a method by which tissue biopsies from patients could be cryo-preserved, stored, or shipped, and later thawed in order to establish live organoid cultures. Methods A method for freezing and recovery, along with straightforward isolation methods using readily available materials were developed. Epithelial isolation and culture conditions were optimized to promote cell survival and the establishment of long-term culture post-thawing. Results Patient biopsies from stomach, duodenum, ileum, colon and from adenomateous colonic polyps were frozen, subsequently thawed and used to successfully establish patient-specific epithelium-only organoid cultures with 100% efficiency (n=31 independent patient biopsies from different regions of the GI tract). Frozen tissue could be shipped internationally and across the United States and used to establish successful organoid cultures. Organoid cultures could be expanded, passaged and frozen down for long-term storage. RNA-sequencing demonstrates that organoids derived from fresh tissue or from frozen biopsies are >99% transcriptionally identical. Conclusions Cryo-preservation of human tissue biopsies followed by the establishment of long-term, expandable cultures has negligible effect on transcription when compared to freshly prepared organoids, and will be of immense benefit to research and for clinical applications. This technique will allow for the establishment of biobanks of human tissues that can be revived and cultured in the future as well as transported across the globe for research, therapeutic or diagnostic use in remote labs and/or clinics.

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Transcriptome analysis of interleukin-4 induced intestinal organoids from Japanese monkey

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