The Generation of Definitive Endoderm from Human Embryonic Stem Cells on 3D Biodegradable Poly(lactic-co-glycolic Acid) Scaffolds and its Comparison to those Generated on 2D Monolayer Cultures

Abstract

Abstract:The generation of insulin producing cells from human embryonic stem cells (hESCs) has shown great promise as a cellular replacement therapy for the treatment of Type 1 Diabetes. Mature functional β-cell surrogates however, have yet to be successfully generated in vivo. One approach to potentially improve current differentiation protocols is the use of 3 dimensional (3D) scaffolds, which has been shown to enhance cellular function and differentiation potential. The present study aimed to explore the feasibility of using single cell preparations of pluripotent hESCs seeded onto laminin or Matrigel coated 3D poly(lactic-co-glycolic) acid (PLGA) scaffolds to derive definitive endoderm, the first vital stage of endoderm tissue differentiation. Our results demonstrated that hESCs which were induced to differentiate on laminin or Matrigel coated 3D scaffolds can be successfully coaxed to differentiate into definitive endoderm. The cells that were cultured on laminin or Matrigel coated 3D scaffolds expressed significantly higher levels of the key endoderm transcription factors SOX17 and FOXA2 in comparison to those differentiated on 2D monolayers. On Matrigel coated 3D scaffolds, the differentiated cells expressed lower levels of the endoderm surface marker CXCR4 and anterior endoderm marker CER in comparison to its monolayer counterpart. Together, the results of this study demonstrated the positive effect of 3D cultures on endoderm commitment from hESCs over traditional monolayer cultures. Furthermore, the definitive endoderm produced on Matrigel coated scaffolds may have a more posterior phenotype in comparison to those derived from monolayers. This may have an effect on later stages of pancreatic differentiation and warrants further detailed investigations.

Enhanced Survival and Function of Islet-Like Clusters Differentiated from Adipose Stem Cells on a Three-Dimensional Natural Polymeric Scaffold: AnIn VitroStudy

“…The macroporous nature of the scaffold provides a spatial environment for maintaining the morphology, thereby sustaining the survival and function of ILCs. Recently, the positive impact of biodegradable PLGA scaffold coated with matrigel on endoderm commitment from human embryonic stem cells over traditional monolayer cultures 48 and reversal of hyperglycemia in diabetic mice using islets derived from embryonic stem cells on PLGA scaffolds 49 were reported. To our knowledge, this is the first report demonstrating the enhanced insulin production of ILCs derived from adipose stem cells on a 3D scaffold.…”

Definitive endoderm differentiation of human-induced pluripotent stem cells using signaling molecules and IDE1 in three-dimensional polymer scaffold

“…[31][32][33] The use of tissue engineering has been actively exploited in recent studies. For example, the successful applications of tissue engineering in the generation of various tissues including bone, 34,35 neural cells, cardiomyocytes, 36,37 hepatocytes, [38][39][40][41] and DE cells 18,29 has attracted a lot of attention in the past decade. 42 In recent years, implementation of synthetic polymers for preparation of scaffolds have dramatically been increased due to their excellent mechanical properties, convenient fabrication processes and also cost effectiveness.…”

“…8,16 In addition to the type of specialized cell per se, the interaction of cells with the extracellular matrix (ECM) plays a central role in controlling cellular behavior, morphology, adhesion, proliferation, migration, and differentiation. 3,4,[17][18][19] Tissue engineering is a newly rapid growing field that aims at developing biological substitutes by incorporating cells or growth factors into the three-dimensional (3D) scaffold, mimicking the native tissue architecture and function. 20,21 These 3D nanofibrous scaffolds are feasibly available by electrospinning method.…”

Delivery of Dissociated Islets Cells within Microporous Annealed Particle Scaffold to Treat Type 1 Diabetes

“…These pseudo‐islet clusters can be formed in microwells, [ 18 ] Matrigel, [ 19 ] or via hanging drop method, [ 20 ] and typically require a 3–5 day “pre‐conditioning” period in vitro prior to implantation. Another alternative delivery method is via synthetic microporous scaffolds, [ 17,20–22 ] which allows for cell self‐organization within pores that can mimic native islet structure and facilitate cell–cell signaling.…”