Controlled induction of human pancreatic progenitors produces functional beta‐like cells in vitro

Abstract

Directed differentiation of human pluripotent stem cells into functional insulin-producing beta-like cells holds great promise for cell replacement therapy for patients suffering from diabetes. This approach also offers the unique opportunity to study otherwise inaccessible aspects of human beta cell development and function in vitro. Here, we show that current pancreatic progenitor differentiation protocols promote precocious endocrine commitment, ultimately resulting in the generation of non-functional polyhormonal cells. Omission of commonly used BMP inhibitors during pancreatic specification prevents precocious endocrine formation while treatment with retinoic acid followed by combined EGF/KGF efficiently generates both PDX1 + and subsequent PDX1 + /NKX6.1 + pancreatic progenitor populations, respectively. Precise temporal activation of endocrine differentiation in PDX1 + /NKX6.1 + progenitors produces glucose-responsive beta-like cells in vitro that exhibit key features of bona fide human beta cells, remain functional after short-term transplantation, and reduce blood glucose levels in diabetic mice. Thus, our simplified and scalable system accurately recapitulates key steps of human pancreas development and provides a fast and reproducible supply of functional human beta-like cells.

MicroRNAs: An adaptive mechanism in the pancreatic β-cell…and beyond?

“…To bring additional clarity to this picture, future studies should address the global temporal regulation and stability of β-cell transcripts in response to changes in extracellular glucose concentrations. The crosslinking protocols that have been developed to identify target sites of miRNAs and RNA-binding proteins can be implemented on the stem cell and fibroblast models now available to understand how non-coding RNAs function in as cells differentiate into insulin-producing β-cells [47] [48] [49] [50]. Improving these protocols to optimize the functional properties and viability of these β-cell models remain a top priority for the development of future therapeutic strategies for combating diabetes.…”

Virgin Beta Cells Persist throughout Life at a Neogenic Niche within Pancreatic Islets

“…This includes the generation of functionally mature human beta cells from stem cells to alleviate the pressing need for an unlimited source of human beta cells to cure T1D -a feat that despite impressive recent progress in the field (Pagliuca et al, 2014;Rezania et al, 2014;Russ et al, 2015) -has not been achieved. We envision that the signaling responses and epigenetic mechanisms that underlie beta cell maturation at the niche overlap with those that are impaired when beta cells dedifferentiate in diabetes.…”

A Novel Pre-Vascularized Subcutaneous Site Safely Accommodates Stem Cell Derived Therapies for Treating Diabetes

“…Positive stimulated C-peptide was found in 12 of 25 (48%) mice in the diabetic group vs. 9 of 25 (36%), in non-diabetic (p = 0. 19). Mean stimulated C-peptide concentrations at 22 weeks were higher in the diabetic group although differences were not statistically significant (0.32 ± 0.15 ng/mL vs. 0.13 ± 0.09 ng/mL, p = 0.30) (Figure 2A).…”

“…Many authors agree on the multiple hurdles these cells encounter in the process of maturation and only recently, successful in vivo maturation have been reported with adequate glucose-response and occasionally, diabetes reversal [15][16][17][18][19].…”