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Generation of mesenchyme free intestinal organoids from human induced pluripotent stem cells
- Source :
- Nature Communications, Nature Communications, Vol 11, Iss 1, Pp 1-15 (2020)
- Publication Year :
- 2020
- Publisher :
- Springer Science and Business Media LLC, 2020.
-
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.<br />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.
- 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
Subjects
Details
- ISSN :
- 20411723
- Volume :
- 11
- Database :
- OpenAIRE
- Journal :
- Nature Communications
- Accession number :
- edsair.doi.dedup.....edb880bb2d5f5325460f600f2654102b