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Title: | ENGINEERING NOVEL SURFACES FOR THE LONG-TERM PROPAGATION OF HUMAN PLURIPOTENT STEM CELLS. | Authors: | YAP YAN WEN | Keywords: | Vitronectin, stem cells, heparan sulfate, pluripotent | Issue Date: | 21-Aug-2012 | Citation: | YAP YAN WEN (2012-08-21). ENGINEERING NOVEL SURFACES FOR THE LONG-TERM PROPAGATION OF HUMAN PLURIPOTENT STEM CELLS.. ScholarBank@NUS Repository. | Abstract: | Defined substrate that can maintain pluripotent stem cells long term is critical for its future regenerative medicine applications. This thesis aims to engineer a tuned heparan sulfate (HS) substrate that is able to bind and present sufficient vitronectin (VN) for the long-term propagation of human embryonic stem cells (hESCs). Results showed that passive adsorption of VN onto tissue-culture treated polystyrene surfaces is inefficient (250 ng/cm2). Therefore by using affinity chromatography technique, this thesis isolated a VN-binding fraction of HS that is able to be immobilized onto allylamine polymerized TCPS by electrostatic interactions which in turn bind and present sufficient VN on surface (150 ng/cm2). The hESCs are able to proliferate on this engineered substrate and continued to retain its pluripotency (morphology, genetic stability, differentiation) for more than 8 passages. This lower VN surface density required on the HS substrate has resulted in a significant savings of ~ 30 %. | URI: | http://scholarbank.nus.edu.sg/handle/10635/47647 |
Appears in Collections: | Ph.D Theses (Open) |
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