Developmentally-Inspired Biomimetic Culture Models to Produce Functional Islet-Like Cells From Pluripotent Precursors

Abstract

Insulin-producing beta cells sourced from pluripotent stem cells hold great potential as a virtually unlimited cell source to treat diabetes. Directed pancreatic differentiation protocols aim to mimic various stimuli present during embryonic development through sequential changes of in vitro culture conditions. This is commonly accomplished by the timed addition of soluble signaling factors, in conjunction with cell-handling steps such as the formation of 3D cell aggregates. Interestingly, when stem cells at the pancreatic progenitor stage are transplanted, they form functional insulinproducing cells, suggesting that in vivo microenvironmental cues promote beta cell specification. Among these cues, biophysical stimuli have only recently emerged in the context of optimizing pancreatic differentiation protocols. This review focuses on studies of cell-microenvironment interactions and their impact on differentiating pancreatic cells when considering cell signaling, cell-cell and cell-ECM interactions. We highlight the development of in vitro cell culture models that allow systematic studies of pancreatic cell mechanobiology in response to extracellular matrix proteins, biomechanical effects, soluble factor modulation of biomechanics, substrate stiffness, fluid flow and topography. Finally, we explore how these new mechanical insights could lead to novel pancreatic differentiation protocols that improve efficiency, maturity, and throughput.

Stepwise differentiation of functional pancreatic β cells from human pluripotent stem cells

“…Alternatively, manipulating the architecture and substrate stiffness of culture platforms to mimic biophysical features of the developmental microenvironment also contributes to pancreatic differentiation (Tran et al, 2020 ; Jiang et al, 2022 ). Air–liquid interface culture (Rezania et al, 2014 ) or suspension culture (Pagliuca et al, 2014 ) allows closer mimicry of in vivo development.…”

Analysis of the transcriptome and metabolome of pancreatic spheroids derived from human induced pluripotent stem cells and matured in an organ-on-a-chip

“…Biomechanical and biochemical stimulations (through shear stress, cell interactions, and soluble factor concentrations) largely impact pancreatic islet function and responses. 38 As microfluidic configuration is critical for islet cultures, 39,40 we calibrated our device with rat islets and iPSC based spheroids in our previous works. Although physiological pancreas islets are exposed to flow rates estimated at 0.03 mL per day (estimation of Sankar et al, 2011 based on the study of Lifson et al, 1985), 40,41 our 20 mL min À1 flow rate led to a nutrient time residence estimated as 100 s. Furthermore, our shear stress of 2 mPa s is in the range of accepted values beneficial for pancreatic in vitro cultures in microfluidic bioreactors (as discussed by Jun et al, 2019).…”

Optimization of differentiation protocols of dental tissues stem cells to pancreatic β-cells

“…However, for initiation of protocol 2, DPSCs & PDLSCs were seeded on freshly coated Geltrex plates acting as an extracellular matrix to enhance stem cells differentiation into beta cells. Extracellular matrix plays a key role in directing stem cells towards differentiation especially in case of pancreatic cell lineage through presenting biochemical signals and growth factors [ 49 ]. Geltrex specifically is a soluble presentation of reduced growth factor basement membrane extract used for support of differentiation cultures.…”