An INSULIN and IAPP dual reporter enables tracking of functional maturation of stem cell-derived insulin producing cells.

ABSTRACT:

Objective

Human embryonic stem cell (hESC; SC)-derived pancreatic β cells can be used to study diabetes pathologies and develop cell replacement therapies. Although current differentiation protocols yield SCβ cells with varying degrees of maturation, these cells still differ from deceased donor human β cells in several respects. We sought to develop a reporter cell line that could be used to dynamically track SCβ cell functional maturation.

Methods

To monitor SCβ cell maturation in vitro, we created an IAPP-2A-mScar and INSULIN-2A-EGFP dual fluorescent reporter (INS^2A-EGFP/+;IAPP^{2A-mScarlet/+}) hESC line using CRISPR/Cas9. Pluripotent SC were then differentiated using a 7-stage protocol to islet-like cells. Immunohistochemistry, flow cytometry, qPCR, GSI

SUBMITTER: Bayly CL 

PROVIDER: S-EPMC11415652 | biostudies-literature | 2024 Nov

REPOSITORIES: biostudies-literature