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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
Author(s) -
McCullough Jeffrey,
Haley Rebecca,
Clay Mary,
Hubel Allison,
Lindgren Bruce,
Moroff Gary
Publication year - 2010
Publication title -
transfusion
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.045
H-Index - 132
eISSN - 1537-2995
pISSN - 0041-1132
DOI - 10.1111/j.1537-2995.2009.02482.x
Subject(s) - cryopreservation , liquid nitrogen , dimethyl sulfoxide , hydroxyethyl starch , apheresis , stem cell , cold storage , nitrogen , andrology , haematopoiesis , materials science , chemistry , biology , biochemistry , immunology , medicine , microbiology and biotechnology , horticulture , platelet , embryo , organic chemistry
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°C mechanical freezer, 3) 5% DMSO and 6% HES with freezing in a −80°C mechanical freezer and storage in a −135°C mechanical freezer, or 4) 5% DMSO and 6% HES with freezing and storage both in a 135°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.