Pancreatic β Cells Derived from Fibroblasts using Non-viral Cell Reprogramming Methods
Loading...
Date
2022-11
Authors
Journal Title
Journal ISSN
Volume Title
Publisher
Abstract
Introduction: Type I diabetes is driven by autoimmune destruction of pancreatic β cells and results in decreased insulin production. Many studies have attempted to generate a functional cell-based therapy to treat type I diabetes, but these approaches have many limitations. To address these limitations, we report on development of a non-viral approach to derive β cells from fibroblasts via electroporation-based gene delivery.
Materials and Methods: We identified 3 transcription factors (3SP) for skin cell plasticity and 7 transcription factors (7βC) for β cell development. Plasmid DNA encoding for these transcription factors or pCMV6 (control plasmid DNA) were delivered to mouse embryonic fibroblasts (MEFs) in vitro using bulk electroporation (BEP). Gene and protein expressions were analyzed at predetermined time points using qRT-PCR or immunohistology as appropriate. Data was collected, processed, and quantified by blinded investigators using Prism GraphPad by Dotmatics and ImageJ.
Results: Fibroblasts transduced with all (n=10) 3SP+7βC factors led to an increase in mRNA expression for both insulin 1 and insulin 2 by day 14 post-BEP (Figure 1A). Moreover, this increased insulin 1/2 expression correlated with a small population (<1%) of transfected cells expressing insulin protein by day 14 post-BEP (Figure 1B).
Conclusion: Our findings suggest that fibroblasts transfected with plasmid DNA encoding for 3SP+7βC factors have the potential to reprogram into induced β cells. Future development of an alternative cell source to treat and potentially cure type I diabetes could greatly improve quality of life for diabetic patients all over the world.
Description
STEP Category: Undergraduate Research
Keywords
Type 1 Diabetes, Non-viral Cell Reprogramming, Gene and Cell Therapy, β Cell Replacement