Date of Graduation

12-2012

Document Type

Thesis

Degree Name

Master of Science in Cell & Molecular Biology (MS)

Degree Level

Graduate

Department

Cell & Molecular Biology

Advisor/Mentor

Jin, Sha

Committee Member

Pinto, Ines

Second Committee Member

Rosenkrans, Charles F. Jr.

Third Committee Member

Wickramasinghe, S. Ranil

Keywords

Biological sciences; Cell biology; Stem cells

Abstract

Human induced pluripotent stem cells (hiPSCs) have the potential to generate patient-specific cells to treat many incurable diseases by cell replacement therapy. However, so far the culture of hiPSCs depends greatly on feeder cells or Matrigel which has safety issues. Thus, chemically defined substrates that could provide niches necessary for cell attachment and proliferation are preferred for clinical application of hiPSCs. Recently, Corning Life Sciences has developed synthetic peptide-functionalized cell culture surface, referred to as Corning® Synthemax that support self-renewal and differentiation of human embryonic stem cell (hESC). In this work, we have collaborated with Corning to investigate the attachment, proliferation, and differentiation of hiPSCs on the Synthemax substrate. We demonstrated that iPS cells retained stable proliferation and pluripotency marker protein expression after growing on the Synthemax substrate for ten consecutive passages. Further examination reveals that integrins áVâ5 mediates attachment to the substrate. Moreover, we observed hiPSCs colonies were more compact on the Synthemax surface. This may be due to less activation of â-catenin-mediated Wnt signaling pathway in cells on the synthetic peptide surface. In hiPSCs grown on the Synthemax Surface, we also found denser actin filaments in the cell-cell interface and down-regulation of vinculin and up-regulation of zyxin, indicating the reorganization of cytoskeleton structure inside cells in response to cell-matrix interaction.

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