Bioengineering microfluidic organoids-on-a-chip

Protocol Exchange
Nikolche GjorevskiOlga Mitrofanova

Abstract

Organoids derived from epithelial stem cells have emerged as powerful platforms to model development and disease in a dish1-3. However, the current mismatch in anatomy, lifespan and size between native organs and their in vitro counterparts severely limits their applicability4. In particular, the closed, cystic architecture of most epithelial stem cell-derived organoids makes experimental manipulation and assay development cumbersome. Here we describe how tissue engineering and cellular self-organization can be combined to guide in vitro organogenesis into openly accessible, functional intestinal tubes termed ‘mini-guts’. Intestinal stem cells (ISCs) rapidly generate simple columnar epithelia when propagated inside basal lamina-like hydrogel scaffolds that feature a tubular and crypt-containing, in vivo-like anatomical structure. Using a microfluidic perfusion system, dead cells shed into the lumen can be continuously removed from the mini-guts. This increases tissue lifespan to months, establishing a homeostatic organoid culture system in which cell proliferation (in crypts) is balanced with cell death (in villus-like domains). The approach developed here can be extended to generate functional tissue/organ models from other ep...Continue Reading

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