1. Generation of mesenchyme free intestinal organoids from human induced pluripotent stem cells
- Author
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Finn Hawkins, Amalia Capilla, Aleksander D. Szymaniak, Alexander Stuffer, Marall Vedaie, Dar Heinze, Aditya Mithal, Carlos Villacorta-Martin, Gustavo Mostoslavsky, Darrell N. Kotton, Megan Peasley, Kristine M. Abo, John Mahoney, Anjali Jacob, and Andrew Berical
- Subjects
0301 basic medicine ,Cystic Fibrosis ,Science ,Mesenchyme ,Cellular differentiation ,Genetic Vectors ,Induced Pluripotent Stem Cells ,Thyroid Nuclear Factor 1 ,Stem-cell differentiation ,General Physics and Astronomy ,Biology ,Cystic fibrosis ,Article ,General Biochemistry, Genetics and Molecular Biology ,Mesoderm ,03 medical and health sciences ,0302 clinical medicine ,Directed differentiation ,Intestine, Small ,medicine ,Organoid ,Humans ,CDX2 Transcription Factor ,Gastrointestinal models ,Gene Knock-In Techniques ,Progenitor cell ,lcsh:Science ,Induced pluripotent stem cell ,CDX2 ,Multidisciplinary ,Disease model ,Cell Differentiation ,Epithelial Cells ,General Chemistry ,medicine.disease ,3. Good health ,Cell biology ,Intestines ,Organoids ,030104 developmental biology ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,lcsh:Q - Abstract
Efficient generation of human induced pluripotent stem cell (hiPSC)-derived human intestinal organoids (HIOs) would facilitate the development of in vitro models for a variety of diseases that affect the gastrointestinal tract, such as inflammatory bowel disease or Cystic Fibrosis. Here, we report a directed differentiation protocol for the generation of mesenchyme-free HIOs that can be primed towards more colonic or proximal intestinal lineages in serum-free defined conditions. Using a CDX2eGFP iPSC knock-in reporter line to track the emergence of hindgut progenitors, we follow the kinetics of CDX2 expression throughout directed differentiation, enabling the purification of intestinal progenitors and robust generation of mesenchyme-free organoids expressing characteristic markers of small intestinal or colonic epithelium. We employ HIOs generated in this way to measure CFTR function using cystic fibrosis patient-derived iPSC lines before and after correction of the CFTR mutation, demonstrating their future potential for disease modeling and therapeutic screening applications., Human induced pluripotent stem cell-derived intestinal organoids (HIOs) are powerful tools to study development and diseases of the gastrointestinal tract. Here, the authors develop a directed differentiation protocol to generate mesenchyme-free HIOs that can be patterned towards proximal small intestine or colonic epithelium, and demonstrated their utility in modeling CFTR function.
- Published
- 2020