Culture of hESC‐derived pancreatic progenitors in alginate‐based scaffolds

Abstract

The effect of alginate-based scaffolds with added basement membrane proteins on the in vitro development of hESC-derived pancreatic progenitors was investigated. Cell clusters were encapsulated in scaffolds containing the basement membrane proteins collagen IV, laminin, fibronectin, or extracellular matrix-derived peptides, and maintained in culture for up to 46 days. The cells remained viable throughout the experiment with no signs of central necrosis.Whereas non-encapsulated cells aggregated into larger clusters, some of which showed signs of morphological changes and tissue organization, the alginate matrix stabilized the cluster size and displayed more homogeneous cell morphologies, allowing culture for long periods of time. For all conditions tested, a stable or declining expression of insulin and PDX1 and an increase in glucagon and somatostatin over time indicated a progressive reduction in beta cellrelated gene expression. Alginate scaffolds can provide a chemically defined, xeno-free and easily scalable alternative for culture of pancreatic progenitors. Although no increase in insulin and PDX1 gene expression after alginate-immobilized cell culture was seen in this study, further optimization of the matrix physicochemical and biological properties and of the medium composition may still be a relevant strategy to promote the stabilization or maturation of stem cell-derived beta cells.

Encapsulation boosts islet-cell signature in differentiating human induced pluripotent stem cells via integrin signalling

“…The number of bihormonal cells was not significantly affected by encapsulation ( Figure 1c). Interestingly, the cells encapsulated during the entire period of differentiation (S7 bead[S0-S7] ) displayed a significant increase of the glucagon-and somatostatin-expressing cell populations, but not insulin, as previously reported 29 .…”

“…In order to eliminate any interference from a possible aggregation/cell clustering effect, we focused on the encapsulation of single cells. This is in contrast with previous studies, which deliberately used clusters to assess the alginate encapsulation effect 29 . We showed here for the first time that The general impact of the 3D organization for the cell differentiation potential is also supported by the introduction of air liquid interface 1 or suspension culture 46 Our subsequent proteome analyses suggested that encapsulation strongly boosts β -cell fate in a stage-specific manner, by promoting early-stage differentiation and potentiating the islet signature during the later stages.…”

“…Microencapsulation of islets into alginate microbeads was used first in the 1980s 21 , and was later employed in several studies for transplantation of pancreatic islets [22][23][24][25] . Previous studies have reported that entrapment of hPSCs under the 3D environment of alginate microcapsules 26 supports long-term maintenance of pluripotency 27 and differentiation of dopamine neurons 28 , as well as pancreatic progenitors 29 . Alginate is recognized for…”

Current and Future Perspectives on Alginate Encapsulated Pancreatic Islet

“…A positive effect of encapsulation has been shown for the development of neonatal porcine islets into mature islets upon culture in serum-containing media [56]. This may be partly due to the hindering of aggregation of clusters upon culture, as has also been shown for human pancreatic progenitors when cultured in alginate beads [57]. hESC have been differentiated to definitive endoderm in alginate capsules [58].…”

“…hESC have been differentiated to definitive endoderm in alginate capsules [58]. For pancreatic progenitors, no effect on differentiation was seen when extracellular matrix (ECM) components (proteins and peptides) were added to the capsule composition [57]. Although no effect was seen in the previous study, the design of the capsule microenvironment to mimic the natural microenvironment has drawn recent attention [59]and positive effects have been shown on beta cell lines [60] as well as on human islets [61].…”