Abstract
BackgroundTo maximize the translational utility of human induced pluripotent stem cells (iPSCs), the ability to precisely modulate the differentiation of iPSCs to target phenotypes is critical. Although the effects of the physical cell niche on stem cell differentiation are well documented, current approaches to direct step-wise differentiation of iPSCs have been typically limited to the optimization of soluble factors. In this regard, we investigated how temporally varied substrate stiffness affects the step-wise differentiation of iPSCs towards various lineages/phenotypes.MethodsElectrospun nanofibrous substrates with different reduced Young’s modulus were utilized to subject cells to different mechanical environments during the differentiation process towards representative phenotypes from each of three germ layer derivatives including motor neuron, pancreatic endoderm, and chondrocyte. Phenotype-specific markers of each lineage/stage were utilized to determine differentiation efficiency by reverse-transcription polymerase chain reaction (RT-PCR) and immunofluorescence imaging for gene and protein expression analysis, respectively.ResultsThe results presented in this proof-of-concept study are the first to systematically demonstrate the significant role of the temporally varied mechanical microenvironment on the differentiation of stem cells. Our results demonstrate that the process of differentiation from pluripotent cells to functional end-phenotypes is mechanoresponsive in a lineage- and differentiation stage-specific manner.ConclusionsLineage/developmental stage-dependent optimization of electrospun substrate stiffness provides a unique opportunity to enhance differentiation efficiency of iPSCs for their facilitated therapeutic applications.
Highlights
To maximize the translational utility of human induced pluripotent stem cells, the ability to precisely modulate the differentiation of Induced pluripotent stem cell (iPSC) to target phenotypes is critical
Our results demonstrate that a sequential application of mechanically distinct electrospun substrates during the differentiation processes can enhance the differentiation efficiency of iPSCs in a lineage- and developmental stage-specific manner
Overall, the results presented in this proof-of-concept study are the first to systematically demonstrate the significant role of the temporally varied mechanical microenvironment on the differentiation of stem cells in a lineage- and developmental stage-specific manner
Summary
To maximize the translational utility of human induced pluripotent stem cells (iPSCs), the ability to precisely modulate the differentiation of iPSCs to target phenotypes is critical. The derivation of human induced pluripotent stem cells (iPSCs) has revolutionized the field of personalized regenerative medicine by offering a potentially unlimited cell source to treat a variety of diseases [1] In addition to their clinical potential, human iPSCs provide opportunities to develop patient-tailored in vitro models for pathogenesis and toxicity studies [2,3,4]. The sequential application of biochemical factors derived from embryonic development provided a foundational backbone to guide iPSC differentiation [10] Such protocols typically discount the role of the underlying physical factors, such as morphology, surface chemistry, and mechanical properties of substrates, which affect differentiation efficiency
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