Generation of gastirc insulin-secreting organoids from human stomach sample.
Xiaofeng HuangQiao ZhouPublished in: Research square (2023)
Stomach stem cells are accessible by biopsy and propagate robustly in culture, offering an invaluable resource for autologous cell therapies. Here we describe a detailed protocol to isolate, expand, engineer and differentiate human gastric stem cells (hGSCs) into pancreatic islet-like organoids containing abundant gastric insulin-secreting (GINS) cells that resemble beta-cells in molecular hallmarks and function. Sequential activation of the inducing factors NGN3 and PDX1-MAFA led hGSCs onto a novel differentiation path, including endocrine progenitor and GINS precursor, before adopting beta-cell identity, at efficiencies close to 70%. GINS organoids acquired glucose-stimulated insulin secretion in 10 days post differentiation.
Keyphrases
- stem cells
- induced pluripotent stem cells
- cell therapy
- induced apoptosis
- type diabetes
- endothelial cells
- cell cycle arrest
- single cell
- randomized controlled trial
- endoplasmic reticulum stress
- oxidative stress
- pluripotent stem cells
- cell death
- bone marrow
- metabolic syndrome
- signaling pathway
- adipose tissue
- platelet rich plasma
- ultrasound guided
- weight loss