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dc.contributor.authorBeatty, Robyneng
dc.contributor.corporatenameUniversity of Missouri-Columbia. Office of Undergraduate Researcheng
dc.contributor.meetingnameUndergraduate Research and Creative Achievements Forum (2006 : University of Missouri--Columbia)eng
dc.date2006eng
dc.date.issued2006eng
dc.descriptionAbstract only availableeng
dc.description.abstractThe transplantation of neuralized stem cells holds the promise of providing neurotrophic effects to individuals suffering from some neurodegenerative disorders. Several current neuralization protocols require the formation of embryoid bodies to obtain a population of neural stem cells. Embryoid bodies (EBs) are round, free-floating aggregates of a heterogeneous population of ES cells undergoing differentiation. The addition of neuralization factors, such as retinoic acid (RA), induces the EB cells toward a neural fate. However, the EBs are only useful if the cells they consist of can survive dissociation. One way of forming EBs is to trypsinize cultured ES cells and transfer them to an uncoated plate with Embryonic Stem cell Induction Media. Different protocols have suggested the use of various trypsin concentrations, ranging from 0.05% to 0.25%, for varying lengths of time. Our current lab protocol uses 0.25% trypsin for 5 minutes, but on occasion this leads to extensive loss of cells during EB dissociation. It was hypothesized that using the high trypsin concentration for an extended period of time was causing the eventual death of the neuralized cells. The goal of my experiments was to optimize the trypsinization protocol that precedes embryoid body formation and to obtain the highest yield of large EBs with the maximum amount of cell survivorship following EB dissociation. Trypsinization conditions that were analyzed included 0.05% trypsin for 5 minutes, 0.25% trypsin for 5 minutes, and 0.25% trypsin for 2.5 minutes. Our results suggest that both 0.25% trypsin for 2.5 minutes and 0.05% trypsin for 5 minutes result in a higher cell survivorship after EB dissociation than the current trypsinization protocol. Cell survivorship was determined by comparing cell counts prior to trypsinization to cell counts following EB dissociation. Future studies could include testing low oxygen tensions and different temperatures during trypsinization.eng
dc.description.sponsorshipLife Sciences Undergraduate Research Opportunity Programeng
dc.identifier.urihttp://hdl.handle.net/10355/899eng
dc.publisherUniversity of Missouri - Columbia Office of Undergraduate Researcheng
dc.relation.ispartofcommunityUniversity of Missouri-Columbia. Office of Undergraduate Research. Undergraduate Research and Creative Achievements Forumeng
dc.subjectneuralized stem cellseng
dc.subjectneurotrophic effecteng
dc.subjectEmbryoid bodies (EBs)eng
dc.subjectneurodegenerative disorderseng
dc.titleOptimization of protocols to propagate and passage undifferentiated and differentiated mouse embryonic stem cells [abstract]eng
dc.typePresentationeng


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