Alginate Encapsulation of Human Embryonic Stem Cells to Enhance Directed Differentiation to Pancreatic Islet-Like Cells

Abstract

The pluripotent property of human embryonic stem cells (hESCs) makes them attractive for treatment of degenerative diseases such as diabetes. We have developed a stage-wise directed differentiation protocol to produce alginate-encapsulated islet-like cells derived from hESCs, which can be directly implanted for diabetes therapy. The advantage of alginate encapsulation lies in its capability to immunoisolate, along with the added possibility of scalable culture. We have evaluated the possibility of encapsulating hESCs at different stages of differentiation. Encapsulation of predifferentiated cells resulted in insufficient cellular yield and differentiation. On the other hand, encapsulation of undifferentiated hESCs followed by differentiation induction upon encapsulation resulted in the highest viability and differentiation. More striking was that alginate encapsulation resulted in a much stronger differentiation compared to parallel two-dimensional cultures, resulting in 20-fold increase in c-peptide protein synthesis. To elucidate the mechanism contributing to encapsulation-mediated enhancement in hESC maturation, investigation of the signaling pathways revealed interesting insight. While the phospho-protein levels of all the tested signaling molecules were lower under encapsulation, the ratio of pSMAD/pAKT was significantly higher, indicating a more efficient signal transduction under encapsulation. These results clearly demonstrate that alginate encapsulation of hESCs and differentiation to islet-cell types provides a potentially translatable treatment option for type 1 diabetes.

Generation and manipulation of hydrogel microcapsules by droplet-based microfluidics for mammalian cell culture

“…MSCs can also undergo myogenic differentiation with successful formation of multinucleated myotubes if they are encapsulated in fibrin hydrogel embedded with fast degradable microbeads 184 . In addition, encapsulated ESCs can differentiate into not only hepatocytes or pancreatic islet-like cells in alginate microcapsules 185, 186 , but also beating cardiomyocytes in liquid core-alginate shell microcapsules 79, 187 . These examples demonstrate that hydrogel microcapsules can guide various stem cells into divergent pathways of differentiations in 3D well-defined microniches.…”

Minireview: Directed Differentiation and Encapsulation of Islet β-Cells—Recent Advances and Future Considerations

“…In another study, alginate-encapsulated hES cell-derived islet-or ␤-like cells were investigated for use in diabetes therapy (108). Encapsulation of mature islet-like cells or hES cell-derived differentiated cells resulted in a low yield of viable cells after maturation, whereas the islet-specific maturation of undifferentiated hES cells obtained under encapsulation was significantly stronger than parallel differentiation conducted in the conventional 2 dimensional configuration.…”

Network Analysis Identifies Crosstalk Interactions Governing TGF-β Signaling Dynamics during Endoderm Differentiation of Human Embryonic Stem Cells

“…: 48-614MAG) according to manufacturer's instructions. The detailed protocol for MagPix is described in our previous study [10]. Mean fluorescence intensity (MFI) was measured using the xMAP (Luminex, Madison, WI, USA) instrument.…”

“…Among these commonly used supplementary constituents, inhibition of PI3K pathway has been consistently reported to enhance endoderm differentiation of hESCs in conjunction with Activin induction [9]. The resulting endoderm cells show good potential to transform into pancreatic β-like cells with continued differentiation [2,10].…”