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Proteomic profiling following cryopreservation
Martin Joseph Vogel
ProQuest / UMI
Wednesday, March 22, 2006
Number of Pages:
To provide a better understanding of the biomolecular response of cells following cryopreservation, Bcl-XL and Bax levels were analyzed by western blot and changes in the proteomic profile were followed via surface enhanced laser desorption ionization-time of flight (SELDI-TOF) technology. In this study, human dermal fibroblast cells were cryopreserved in media + 5% DMSO or CryoStor-CS5. Protein was isolated from cells at time points post-thaw. Utilization of CryoStor-CS5 yielded an increase in survival over that of media + 5% DMSO. The Bax/Bcl-XL ratio was elevated following cryopreservation. Utilization of CryoStor-CS5 resulted in a decrease in the pro-anti-apoptotic protein ratio. Protein profile patterns of cryopreserved cells using SELDI-TOF differed from that of controls. Additionally, the protein profile from samples stored in different solutions were similar immediately post-thaw but displayed different profiles at subsequent time-points. Using database searches, possible cryopreservation-induced biomarkers were implicated in survival and/or death of cells following cryopreservation.
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