Membrane and Mitochondrial Responses to Cryobiological Conditions

• Author / Creator

  Reardon, Anthony J

• The success of cellular cryopreservation is indicated by the state of post-thawed cells. Using human umbilical vein endothelial cells as a model, the cryobiological response to interrupted cooling protocols was evaluated with a combination of fluorescence microscopy and flow cytometry. Cryoinjured cells were identified using fluorescence intensity with flow cytometry, where light scatter gating strategies were shown to have limitations. Plasma membrane integrity has become the standard assay for post-thaw cell survival. This investigation provided a direct comparison of the integrity of the plasma membrane with the functional state of mitochondria in frozen-thawed cells. A disconnect was found between these two sites of cryoinjury: at specific low temperature conditions cells with depolarized mitochondria still had an intact plasma membrane. This result indicates that different mechanisms may affect each of these sites of cryoinjury. These findings are of interest in understanding the mechanisms of cryoinjury required to minimize cell damage during cryopreservation.

• Subjects / Keywords

  • Cryobiology
  • Plasma Membrane
  • Flow Cytometry
  • Mitochondria

• Graduation date

  Spring 2013

• Type of Item

  Thesis

• Degree

  Master of Science

• DOI

  https://doi.org/10.7939/R38H9X

• License

  This thesis is made available by the University of Alberta Libraries with permission of the copyright owner solely for non-commercial purposes. This thesis, or any portion thereof, may not otherwise be copied or reproduced without the written consent of the copyright owner, except to the extent permitted by Canadian copyright law.