Effect of substrate stiffness on early human embryonic stem cell differentiation

Journal of Biological Engineering

Raj R Rao²  Vamsi K Yadavalli²  Rukmani Ramachandran²  Nikolai Eroshenko¹ 

[1] Current Address: School of Engineering and Applied Sciences, Harvard University, Boston, MA, USA;Department of Chemical and Life Science Engineering, Virginia Commonwealth University, Richmond, VA, USA

关键词: Stiffness;    Differentiation;    Extracellular matrix;    Biomaterials;    Stem cells;   

Others  :  805917
DOI  :  10.1186/1754-1611-7-7

 received in 2012-04-11, accepted in 2013-03-14,  发布年份 2013

【 摘 要 】

Background

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. 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.

Results

PDMS 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.

Conclusions

We 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.

【 授权许可】

   
2013 Eroshenko et al.; licensee BioMed Central Ltd.

【 图 表 】

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