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Cryopreservation and Hypothermal Storage of Hematopoietic Stem Cells

AlMulhem, Norah
http://orcid.org/0000-0002-4247-0399
http://rave.ohiolink.edu/etdc/view?acc_num=ucin1439307244

Abstract Details

2015, MS, University of Cincinnati, Allied Health Sciences: Transfusion and Transplantation Sciences.
The recent availability of commercially available storage media (CryoStor™ and HypoThermosol™) designed for optimal long-term and short-term hematopoietic stem cell (HSC) storage prompted an evaluation of hematopoietic stem cell and hematopoietic progenitor cell (HSC/P) viability and functionality after storage in these media formulations, compared with the conventional media used at Hoxworth Blood Center. Three human umbilical cord blood units (CBUs) were cryopreserved in CryoStor5 (CS5), CryoStor10 (CS10), and a conventional internally prepared cryopreservation medium, then analyzed post-thaw for viability and recovery of several mature and immature hematopoietic cell types, as well as for clonogenic capacity, and proliferation potential. Flow cytometric analysis indicated similar post-thaw viability of most cell subsets cryopreserved in CS5, and CS10 compared to the conventional cryopreservation medium (containing 5 % Dimethylsulfoxide (DMSO) and 2.5 % hydroxyethyl starch). This variation in viability was not statistically significant (p-value 0.2-1). Bromodeoxyuridine (BrdU) uptake was used to measure the ability of the frozen/thawed cells to proliferate in culture for 48 h in response to stem cell factor (SCF), FLt-3 ligand (FLt-3) and thrombopoietin (TPO). Proliferation potential and clonogenic capacity were both slightly better with after freezing in CS10; however, the differences were not statistically significant. This study shows that the conventional medium for cryopreservation used in our laboratory is similarly effective, compared with CS5 or CS10 media, in protecting the cryopreserved CBU derived HSC/P products. The same analytical methods were used to compare HypoThermosol® (HTR-FRS®), which is designed for short-term refrigerated storage of hematopoietic cells, to a locally prepared medium containing Plasma-Lyte A and 0.5 % human serum albumin (HSA). Measurements were performed after 24, 48 and 72 h of cold storage (4°C). Results showed similar viability and recovery after 24 h of storage, but after 48 and 72 h, a significant decline in viability occurred in a few of the subsets when stored in Plasma-Lyte A/HSA medium, compared to when stored in HTS-FRS®. Differences in clonogenic capacity and proliferation potential were not significant, however the cells’ proliferation potential was slightly better after storage in HTS-FSR®. Taken together, these results indicate that the HTS-FRS® storage media preserves hematopoietic cell function better than Plasma-Lyte A/ 0.5 % HSA, especially if the cells are to be stored for more than 24 hours. It is possible that these in-vitro results could translate to improved engraftment after storing umbilical cord blood, bone marrow or mobilized peripheral blood in these new media.
Thomas Leemhuis, Ph.D. (Committee Chair)
Jose Cancelas-Perez, M.D. (Committee Member)
Patricia Morgan Carey, M.D. (Committee Member)
Carolyn Lutzko, Ph.D. (Committee Member)
70 p.
Health Sciences
Cryopreservation; Hypothermal storage; HPC P; CryoStor; HypoThermosole

Recommended Citations

Citations

  • AlMulhem, N. (2015). Cryopreservation and Hypothermal Storage of Hematopoietic Stem Cells [Master's thesis, University of Cincinnati]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1439307244

    APA Style (7th edition)

  • AlMulhem, Norah. Cryopreservation and Hypothermal Storage of Hematopoietic Stem Cells. 2015. University of Cincinnati, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=ucin1439307244.

    MLA Style (8th edition)

  • AlMulhem, Norah. "Cryopreservation and Hypothermal Storage of Hematopoietic Stem Cells." Master's thesis, University of Cincinnati, 2015. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1439307244

    Chicago Manual of Style (17th edition)

ucin1439307244
4,173
© 2015, some rights reserved.Creative Commons License Cryopreservation and Hypothermal Storage of Hematopoietic Stem Cells by Norah AlMulhem is licensed under a Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. Based on a work at etd.ohiolink.edu.
This open access ETD is published by University of Cincinnati and OhioLINK.