Combinatorial Extracellular Matrices for Human Embryonic Stem Cell Differentiation in 3D

David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, 45 Carleton Street, E25-342, Cambridge, Massachusetts 02139, USA.
Biomacromolecules (Impact Factor: 5.75). 08/2010; 11(8):1909-14. DOI: 10.1021/bm100357t
Source: PubMed

ABSTRACT Embryonic stem cells (ESCs) are promising cell sources for tissue engineering and regenerative medicine. Scaffolds for ESC-based tissue regeneration should provide not only structural support, but also signals capable of supporting appropriate cell differentiation and tissue development. Extracellular matrix (ECM) is a key component of the stem cell niche in vivo and can influence stem cell fate via mediating cell attachment and migration, presenting chemical and physical cues, as well as binding soluble factors. Here we investigated the effects of combinatorial extracellular matrix proteins on controlled human ESC (hESC) differentiation. Varying ECM compositions in 3D markedly affects cell behavior, and optimal compositions of ECM hydrogels are identified that facilitate specific-lineage differentiation of stem cells. To our knowledge, this is the first combinatorial analysis of ECM hydrogels for their effects on hESC differentiation in 3D. The 3D matrices described herein may provide a useful platform for studying the interactive ECM signaling in influencing stem cell differentiation.

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Available from: Lily Keung, May 12, 2014
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    • "However, these biocompatible materials do not provide appropriate ECM-mediated biological signals for enhanced cellular attachment and migration. Thus, the presentation of appropriate chemical and physical cues onto nanofibers are required for enhanced cellular attachment and bioactivity [28]. Fibronectin (FN) is known as an important ECM molecule for stem cell adhesion, survival and differentiation [29] [30] [31]. "
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    ABSTRACT: Stem cells seeded onto biofunctional materials have greater potency for therapeutic applications. We investigated whether umbilical cord blood-derived mesenchymal stem cell (UCB-MSC)-seeded fibronectin (FN)-immobilized polycaprolactone (PCL) nanofibers could improve cardiac function and inhibit left ventricle (LV) remodeling in a rat model of myocardial infarction (MI). Aligned nanofibers were uniformly coated with poly (glycidyl methacrylate) by initiated chemical vapor deposition followed by covalent immobilization of FN proteins. Degree of cell elongation and adhesion efficacy was improved by FN immobilization. Furthermore, genes related to angiogenesis and mesenchymal differentiations were up-regulated in the FN-immobilized PCL nanofibers in comparison to control PCL nanofibers in vitro. Four weeks after the transplantation in the rat MI model, the echocardiogram showed that the UCB-MSC-seeded FN-immobilized PCL nanofiber group increased LV ejection fraction and fraction shortening as compared to the non-treated control and acellular FN-immobilized PCL nanofiber groups. Histological analysis indicated that the implantation of UCB-MSCs with FN-immobilized PCL nanofiber induced a decrease in MI size and fibrosis, and an increase in scar thickness. This study indicates that FN-immobilized biofunctional PCL nanofibers could be an effective carrier for UCB-MSC transplantation for the treatment of MI.
    Acta biomaterialia 03/2014; 10(7). DOI:10.1016/j.actbio.2014.03.013 · 5.68 Impact Factor
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    • "Multiwell-based assays have been well accepted for screening of stem cell fate inside combinatorial environments7131415. These, platforms are typically generated by adding stem cell encapsulated hydrogels and ECM proteins into standard multiwell plates13. Other approaches have relied on the deposition and freeze drying of polymeric solutions into multiwell plates to generate scaffold libraries capable of screening cell-matrix interactions within 3D microenvironments1415. "
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    ABSTRACT: Development of three dimensional (3D) microenvironments that direct stem cell differentiation into functional cell types remains a major challenge in the field of regenerative medicine. Here, we describe a new platform to address this challenge by utilizing a robotic microarray spotter for testing stem cell fates inside various miniaturized cell-laden gels in a systematic manner. To demonstrate the feasibility of our platform, we evaluated the osteogenic differentiation of human mesenchymal stem cells (hMSCs) within combinatorial 3D niches. We were able to identify specific combinations, that enhanced the expression of osteogenic markers. Notably, these 'hit' combinations directed hMSCs to form mineralized tissue when conditions were translated to 3D macroscale hydrogels, indicating that the miniaturization of the experimental system did not alter stem cell fate. Overall, our findings confirmed that the 3D cell-laden gel microarray can be used for screening of different conditions in a rapid, cost-effective, and multiplexed manner for a broad range of tissue engineering applications.
    Scientific Reports 01/2014; 4:3896. DOI:10.1038/srep03896 · 5.58 Impact Factor
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    • "Moreover, instead of the common growth factor-induced differentiation system on tissue culture plates (TCPs) in vitro, various biomaterials are constructed as two-or three-dimensional (2D or 3D) matrices to help to guide stem cells differentiation into specific cell types (Haque et al., 2012; Li et al., 2011). The mechanical properties (Engler et al., 2006), chemical composition (Yang et al., 2010), structure and architecture (Xie et al., 2009) of scaffolds can all contribute to modulating the differentiation of stem cells. However, so far, few reports have revealed the underlying mechanisms by which scaffolds modulate the differentiation of stem cells (Park et al., 2011; Keselowsky et al., 2005). "
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    ABSTRACT: Different hydrogel materials have been prepared to investigate the effects of culture substrate on the behaviour of pluripotent cells. In particular, genipin-crosslinked gelatin–silk fibroin hydrogels of different compositions have been prepared, physically characterized and used as substrates for the culture of pluripotent cells. Pluripotent cells cultured on hydrogels remained viable and proliferated. Gelatin and silk fibroin promoted the proliferation of cells in the short and long term, respectively. Moreover, cells cultured on genipin-crosslinked gelatin–silk fibroin blended hydrogels were induced to an epithelial ectodermal differentiation fate, instead of the neural ectodermal fate obtained by culturing on tissue culture plates. This work confirms that specific culture substrates can be used to modulate the behaviour of pluripotent cells and that our genipin-crosslinked gelatin–silk fibroin blended hydrogels can induce pluripotent cells differentiation to an epithelial ectodermal fate.
    Journal of Tissue Engineering and Regenerative Medicine 01/2014; DOI:10.1002/term.1868 · 4.43 Impact Factor
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