alexa Long-term storage of peripheral blood stem cells frozen and stored with a conventional liquid nitrogen technique compared with cells frozen and stored in a mechanical freezer.


Journal of Blood Disorders & Transfusion

Author(s): McCullough J, Haley R, Clay M, Hubel A, Lindgren B,

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Abstract BACKGROUND: Cryopreservation of hematopoietic progenitor cells using liquid nitrogen and controlled-rate freezing requires complex equipment and highly trained staff and is expensive. We compared the liquid nitrogen method with methods using a combination of dimethyl sulfoxide (DMSO) and hydroxyethyl starch (HES) for cryopreservation followed by storage in mechanical freezers. STUDY DESIGN AND METHODS: Peripheral blood stem cells (PBSCs) were collected from normal donors by apheresis and allocated to one of four preservation and storage conditions: 1) 10\% DMSO with freezing in liquid nitrogen and storage in liquid nitrogen, 2) 5\% DMSO and 6\% HES with freezing and storage in a -80 degrees C mechanical freezer, 3) 5\% DMSO and 6\% HES with freezing in a -80 degrees C mechanical freezer and storage in a -135 degrees C mechanical freezer, or 4) 5\% DMSO and 6\% HES with freezing and storage both in a 135 degrees C mechanical freezer. Cells were stored for 5 years during which total nucleated cells (TNCs), cell viability, CD34+ cell content, and colony-forming unit-granulocyte-macrophage content were determined. RESULTS: There were some significant differences in the variables measured during freezing and the 5 years of storage compared to the values before freezing and storage; however, these differences were not consistent and do not favor one protocol over the others. Samples stored for 24 hours before cryopreservation showed a significant decrease in TNCs, but no other significant changes during the 5 years. CONCLUSION: In vitro measurements indicate that PBSCs can be successfully frozen and stored using a combination of DMSO and HES providing smaller amounts of DMSO and allowing simplified freezing and storage conditions. This article was published in Transfusion and referenced in Journal of Blood Disorders & Transfusion

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