Fabrication of synthetic polymer coatings and their use in feeder-free culture of human embryonic stem cells |
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Authors: | Nandivada Himabindu Villa-Diaz Luis G O'Shea K Sue Smith Gary D Krebsbach Paul H Lahann Joerg |
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Institution: | Department of Chemical Engineering, University of Michigan, Ann Arbor, Michigan, USA. |
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Abstract: | The culture of human embryonic stem (hES) cells in defined and xenogeneic-free conditions will contribute substantially to future biotechnological and medical applications. To achieve this goal, we developed the first fully defined synthetic polymer coating poly2-(methacryloyloxy)ethyl dimethyl-(3-sulfopropyl)ammonium hydroxide] (PMEDSAH) that sustains long-term growth of hES cells in different culture media. Here we describe a detailed protocol for the reproducible fabrication of PMEDSAH coating on tissue culture polystyrene dishes, and for the feeder-free culture of hES cells on PMEDSAH coating in defined culture medium. This culture system represents a key step toward the fully defined and xenogeneic-free culture of hES cells. |
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