2010•UpSpace Institutional Repository (University of Pretoria)Requires access
Flow cytometric analysis of stem cells derived from umbilical cord blood
Doretha Thompson
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Abstract
Successful engraftment is highly dependent on the quality and quantity of stem cells and nucleated cells in cord blood. Storage of umbilical cord blood is expensive and it will be very useful if factors that influence cell count and viability could be identified to aid in the decision to process and store cord blood collections for the ultimate aim of successful engraftment. This study determined the standards for laboratory parameters of haematopoietic potential, such as collection volume, post processing volume, CD34+/45+ cell counts and viability of stem cells and leukocytes and cell ratios for the cord blood bank. In this research we determined whether maternal age and infant gender have an effect on laboratory parameters. We studied the effect of 4°C and room temperature (RT) as well as the period of storage on the same laboratory parameters. The quality and recovery of stem cells and leukocytes after laboratory manipulation was determined. Established standards for leukocyte count, stem cell count and collection volume compare well with other international UCB banks. Maternal age and infant gender have no influence on laboratory parameters and could therefore not be used as a measure of cell quantity and quality prior to processing. Cell count and cell viability of stem cells is not significantly influenced by 4°C or RT temperature or 24h and 48h storage. Leukocyte cell count and viability is not affected by storage temperature, but increased storage periods showed high levels of leukocyte cell count deterioration and decreased leukocyte cell viability. Processing of UCB causes significant cell loss in stem cells and leukocytes. Processing or no processing of UCB has showed no affect on the viability of stem cells stored at different storage periods and temperature. Temperature has no affect on leukocyte cell counts and viability of either processed or unprocessed leukocytes but increased storage periods dramatically decrease leukocyte count and viability. The information generated by this study will assist in the process of optimizing the storage of UCB. Copyright