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Please cite this article as: Best, M., Carroll, M., Hanley, N.A., Hanley, K.P., Embryonic stem cells to beta-cells by understanding pancreas development, Molecular and Cellular Endocrinology (2007), doi:10.1016/j.mce.2008 This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.

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.