Abstract
BackgroundThe pluripotency and self renewing properties of human embryonic stem cells (hESC) make them a valuable tool in the fields of developmental biology, pharmacology and regenerative medicine. Therefore, there exists immense interest in devising strategies for hESC propagation and differentiation. Methods involving simulation of the native stem cell microenvironment, both chemical and physical, have received a lot of attention in recent years. Equally important is evidence that cells can also sense the mechanical properties of their microenvironment. In this study, we test the hypothesis that hESCs accept mechanical cues for differentiation from the substrate by culturing them on flexible polydimethylsiloxane (PDMS) of varying stiffness.ResultsPDMS substrates were prepared using available commercial formulations and characterized for stiffness, surface properties and efficiency of cell attachment and proliferation. Across different substrate stiffness, cell numbers, cell attachment and cell surface area were found to be similar. Expression of pluripotency markers decreased with increased time in culture across all PDMS substrates of varying stiffness. Analysis of gene expression of differentiation markers indicates that the differentiation process becomes less stochastic with longer culture times.ConclusionsWe evaluated the utility of PDMS substrates for stem cell propagation and substrate mediated differentiation. The stiffness affected gene expression of pluripotent and differentiation markers with results indicating that these substrate systems could potentially be used to direct hESC fate towards early mesodermal lineages. This study suggests that coupled with soluble factors, PDMS substrates could potentially be useful in generating defined populations of differentiated cells.
Highlights
The pluripotency and self renewing properties of human embryonic stem cells make them a valuable tool in the fields of developmental biology, pharmacology and regenerative medicine
The substrate acts as the primary extra cellular matrix (ECM) component, and while a feeder layer of inactivated mouse embryonic fibroblasts (MEF) has been the traditional gold standard, polymeric materials have been investigated for their ability to support human embryonic stem cells (hESC) propagation
In this study, we have demonstrated that fibronectincoated PDMS substrates are capable of supporting hESC attachment and proliferation
Summary
The pluripotency and self renewing properties of human embryonic stem cells (hESC) make them a valuable tool in the fields of developmental biology, pharmacology and regenerative medicine. Human embryonic stem cells (hESCs) are characterized by their ability to self renew and to differentiate into any diploid human cell type. This property makes them a valuable tool for studying the basic biology of lineage specification, and for applications in fields such as pharmacology and tissue engineering [1]. The ECM as a major niche element provides a scaffold for cellular support, migration and proliferation, and acts as the surrounding microenvironment that influences the cellular fate decision by presenting physical and chemical cues as well as binding soluble factors [12]. We have previously reviewed the potential of bio-inspired polymers in determining human stem cell fate, which possess the advantage of being a xeno-free culture system but can be tailored to very specific needs [14]
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