Endothelial Cells Mediate Islet-Specific Maturation of Human Embryonic Stem Cell-Derived Pancreatic Progenitor Cells

Abstract

It is well recognized that in vitro differentiation of embryonic stem cells (ESC) can be best achieved by closely recapitulating the in vivo developmental niche. Thus, implementation of directed differentiation strategies has yielded encouraging results in the area of pancreatic islet differentiation. These strategies have concentrated on direct addition of chemical signals, however, other aspect of the developmental niche are yet to be explored. During development, pancreatic progenitor (PP) cells grow as an epithelial sheet, which aggregates with endothelial cells (ECs) during the final stages of maturation. Several findings suggest that the interactions with EC play a role in pancreatic development. In this study, we recapitulated this phenomenon in an in vitro environment by maturing the human ESC (hESC)-derived PP cells in close contact with ECs. We find that co-culture with different ECs (but not fibroblast) alone results in pancreatic islet-specific differentiation of hESC-derived PP cells even in the absence of additional chemical induction. The differentiated cells responded to exogenous glucose levels by enhanced C-peptide synthesis. The co-culture system aligned well with endocrine development as determined by comprehensive analysis of involved signaling pathways. By recapitulating cell-cell interaction aspects of the developmental niche we achieved a differentiation model that aligns closely with islet organogenesis.

Capsule stiffness regulates the efficiency of pancreatic differentiation of human embryonic stem cells

“…Alternately, β cells can be derived from human pluripotent stem cells (hPSCs), which have the capacity to self-renew and differentiate, making them attractive for regenerative medicine and cell therapy applications. A significant amount of work has focused on deriving β -like cells from pluripotent stem cells, including work done in our own lab [3][4][5][6]. Recently, the Melton group and the Kieffer group have reported a significant breakthrough in generating glucose responsive insulin producing β cells from hPSCs in vitro [3,7].…”

Development of an Alginate Array Platform to Decouple the Effect of Multiparametric Perturbations on Human Pluripotent Stem Cells During Pancreatic Differentiation

“…Having confirmed germ layer commitment, we next analyzed pancreatic differentiation by quantifying the expression levels of PDX1 and Nkx6.1 proteins. Both PDX1 and Nkx6.1 are vital markers of the progenitor population required for downstream β cell commitment, and are extensively used to characterize the pancreatic progenitor stage . As illustrated in Figure B, both SC3 and SC5 had comparable levels of PDX1 and Nkx6.1 expression levels, with PDX1 being only slightly higher in SC5 and Nkx6.1 slightly higher in SC3 conditions.…”

“…Encapsulated hESCs were first induced to the DE stage (Figure S1B, Supporting Information), and differentiation was evaluated by SOX17 and FOXA2 protein immunostaining. These are key protein markers typically used to identify successfully DE stage differentiation Figure A presents the protein expression levels quantified using the LICOR Odyssey platform, and normalized to DNA for cell differentiated to the DE stage. Control culture of undifferentiated cells stained for SOX17 and FOXA2 showed minimal expression prior to DE inductions (Figure S2A, Supporting Information).…”

“…These cells have two distinct characteristics which make them highly attractive: they can become any cell type in the body, and can self‐renew indefinitely. Over the last two decades there has been concentrated effort to derive functional organ specific cells from hPSCs, which include, but are not limited to, cardiac cells, neurons, hepatocytes, and pancreatic beta cells . With the recent success in deriving functional cells from hPSCs, the current emphasis is on translating these cells to the clinic, as a regenerative cell source to replace donor organs.…”

Endothelial cell regulation of salivary gland epithelial patterning

“…Multiple angiocrine factors are produced by endothelial cells in every organ, and the profile of angiocrine factors is known to be organ specific (Rafii et al, 2016;Azizoglu and Cleaver, 2016). Angiocrine factors are known to regulate epithelial cell behaviors such as survival and proliferation, polarity, and differentiation (Ingthorsson et al, 2010;Hagiwara et al, 2015;Jaramillo et al, 2015;Kao et al, 2015). Recent evidence demonstrates that vascular endothelial cell-mediated signaling regulates stem/ progenitor cell renewal and differentiation during organ development, homeostasis and repair/regeneration (Rafii et al, 2016).…”