Dielectric spectroscopy for monitoring human pancreatic islet differentiation within cell-seeded scaffolds in a perfusion bioreactor system.
The long-term in vitro culture and differentiation of human pancreatic islets is still hindered by the inability to emulate a suitable microenvironment mimicking physiological extracellular matrix (ECM) support and nutrient/oxygen perfusion. This is further amplified by the current lack of a non-invasive and rapid monitoring system to readily evaluate cellular processes. In this study, we realized a viable method for non-invasively monitoring isolated human pancreatic islets in vitro. Islets are induced to dedifferentiate into proliferative duct-like structures (DLS) in preparation for potential and subsequent re-differentiation into functional islet-like structures (ILS) in a process reminiscent of islet regeneration strategies. This long-term in vitro process is conducted within a three-dimensional microenvironment involving islets embedded in an optimized ECM gel supported by microfabricated three-dimensional scaffolds. The islet-scaffold is then housed and continuously perfused within chambers of a bioreactor platform. The process in its entirety is monitored through dielectric spectroscopy measurements, yielding an accurate representation of cellular morphology, functionality, and volume fraction. This non-invasive and real-time monitoring tool can be further manipulated to elucidate important information about the optimized cellular microenvironment required for maintaining long-term culture and achieve efficient differentiation for islet regeneration.
Biomedical Engineering Department, McGill University, 3775 University Street, Montreal, Quebec H3A 2B4, Canada. maryam.tabrizian@mcgill.ca.
J
J
Daoud
K
K
Heileman
S
S
Shapka
L
L
Rosenberg
M
M
Tabrizian
D001672:Biocompatible Materials / Q000494:pharmacology; D019149:Bioreactors; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D058266:Dielectric Spectroscopy / Q000379:methods*; D005109:Extracellular Matrix / Q000378:metabolism; D020342:Finite Element Analysis; D006801:Humans; D007515:Islets of Langerhans / Q000166:cytology* / Q000187:drug effects; D010100:Oxygen / Q000378:metabolism; D010477:Perfusion; D054457:Tissue Scaffolds
D001672:Biocompatible Materials; D010100:Oxygen