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

Abstract

Human-induced pluripotent stem cells (hiPSCs) are considered to be potentially able to differentiate into all human cell lineages and thus hold promise as an unlimited source for cell replacement therapies in clinical applications. Definitive endoderm (DE) formation is the first and crucial step in the development of visceral organs such as liver, lung, pancreas and so forth. Therefore, efficient generation of DE cells ensures the efficient generation of eventual target cells used in cell therapy. In the present study, Matrigel-coated poly(lactic acid)/gelatin (PLA/gelatin) nanofibrous scaffolds were utilized to investigate the proliferation and differentiation of hiPSCs into DE cells. Analyses of DE-specific markers including Sox17, FoxA2, and Gooscoid (Gsc) genes revealed higher levels of mRNA and protein expression in the differentiated hiPSCs cells cultured on PLA/gelatin scaffolds than cells differentiated in two-dimensional (2D) culture. Our results showed that three-dimensional (3D) cultures could significantly promote DE differentiation in comparison with 2D culture. Also using small molecules such as inducer of definitive endoderm 1 (IDE1) and signaling molecules such as Activin A and Wnt3a could enhance the DE differentiation of hiPSCs with Activin A/Wnt3a being significantly more potent in both 2D and 3D cultures compared to IDE1. The results of this study may have impact in tissue engineering and cell replacement therapy of visceral organs-related diseases.

Generation of insulin‐producing cells from human adipose‐derived mesenchymal stem cells on PVA scaffold by optimized differentiation protocol

“…In comparison to 2D culture systems, 3D culture improves islet like morphology, differentiation, and function. These results correspond with the previous findings of the studies on the types of stem cells (Aloysious & Nair, ; Hoveizi et al, ; Khorsandi, Khodadadi, Nejad‐Dehbashi, & Saremy, ). The role of hADSCs in tissue engineering and regenerative medicine is promising; however, bioartificial pancreas based on tissue engineering still appears to be fantastic.…”

In vitro comparative survey of cell adhesion and proliferation of human induced pluripotent stem cells on surfaces of polymeric electrospun nanofibrous and solution‐cast film scaffolds

“…19,20 Electrospun nanofibrous scaffolds possess many unique properties, including extremely high surface-to-volume ratio, appropriate porosity, and malleability to conform to a wide variety of sizes and shapes which are similar to the ECM and can enhance the adhesion, proliferation and growth of cells. 2,4 Previous findings from many other researchers were consistent with ours, which also demonstrated that the nanoarchitecture of the scaffold has significant effects on various cell behaviors including cell adhesion, orientation and proliferation. [21][22][23] Here, although we found that hiPSCs can be cultured on electrospun and film scaffolds but we repeatedly found significantly better performance of hiPSCs on electrospun scaffold over film in our adhesion, viability and proliferation assays.…”

“…Nanofibrous scaffolds produced by electrospinning have a great 12 potential in many biomedical applications and they are suitable substrates to promote cell attachment, proliferation, and differentiation 19,20 . Electrospun nanofibrous scaffolds possess many unique properties, including extremely high surface-to-volume ratio, appropriate porosity, and malleability to conform to a wide variety of sizes and shapes which are similar to the ECM and can enhance the adhesion, proliferation and growth of cells 2,4 . Previous findings from many other researchers were consistent with ours, which also demonstrated that the nanoarchitecture of the scaffold has significant effects on various cell behaviors including cell adhesion, orientation and proliferation [21][22][23] .…”

“…14 In recent years the utility of different biomaterials for regenerative medicine applications was investigated. 2 Tissue Engineering has been developed to design suitable biomaterials to substitute irreversibly damaged tissues and organs. Tissue engineering is a new approach in which techniques are being developed to transplant autologous cells onto suitable scaffolds to eventually form new functional tissues.…”

“…Experimental manipulation of hiPSCs differentiation provides a model of mammalian development that is amenable to molecular and cellular analysis. 2 ECM components play a critical role in regulating stem cell mobilization. ECM signaling and interactions between cells and the ECM are mainly mediated by the integrin family of cell-cell and cell-matrix receptors.…”