Optimization of Murine Pancreas Decellularization to Obtain ECM for a Co-Culture System Capable of Producing Mature Pancreas Cell Types
Year: 2020
Type: dissertation
Abstract: Maturation and cell organization have remained issues for in vitro pancreatic tissue engineering. However, many culture conditions do not incorporate the extracellular matrix (ECM), an element crucial to organogenesis. This project aimed to create a 3D co-culture system with murine ECM. To obtain ECM, optimization of decellularization was explored using perfusion of whole pancreas or lobe as well as by using soak-agitation for slices. The detergents Triton X-100, CHAPS and SDS were tested. ECMs were analyzed for DNA content and retention of critical ECM proteins. CHAPS was chosen for further optimization with benzonase and resulted in decellularized ECM that retained key proteins. This ECM was tested as both a 2D coating and a 3D intact tissue slice matrix for survival and maturation of embryonic stem cells. Only the 3D slices suggested maturation toward a pancreatic lineage. Therefore, this study created a 3D co-culture system to help further tissue engineering efforts.
Source: TSpace (University of Toronto)
Author: Amanda Fantin
Language: English
Subfield: Surgery
Field: Medicine
Domain: Health Sciences
Open Access status: green
OpenAlex ID: W3183443153