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Prolonged cryopreservation of human bone marrow

L M Parker, N Binder, R Gelman

    Transplantation
    |June 1, 1981
    PubMed
    Summary

    Human bone marrow cryopreservation using vapor phase liquid nitrogen storage for over 40 months is viable. This method effectively preserves nucleated cells and granulocyte-monocyte colony-forming cells (CFU-c) for transplantation.

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    Area of Science:

    • Hematology
    • Cryobiology
    • Cellular Biology

    Background:

    • Effective cryopreservation of human bone marrow is crucial for hematopoietic stem cell transplantation.
    • Long-term storage viability of bone marrow cells, particularly hematopoietic stem cells, needs rigorous assessment.

    Purpose of the Study:

    • To evaluate the viability of human bone marrow after prolonged storage in the vapor phase of liquid nitrogen.
    • To assess the impact of cryopreservation and thawing on nucleated cell count and colony-forming cell (CFU-c) efficiency.

    Main Methods:

    • Human bone marrow from eight patients was cryopreserved for 40-42 months in 10% DMSO.
    • Cell counts and CFU-c plating efficiency were measured before freezing, after thawing, and after DMSO removal and incubation.
    • Discontinuous centrifugation was used to concentrate nucleated cells.

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    Main Results:

    • Prolonged vapor phase storage resulted in a median cell loss of 9% and a 2% increase in CFU-c plating efficiency.
    • DMSO dilution, washing, and incubation led to cell losses of 19-24% but increased CFU-c plating efficiency by 22-79%.
    • Viable CFU-c counts remained comparable to pre-storage levels throughout the process.

    Conclusions:

    • Vapor phase liquid nitrogen storage is a suitable method for long-term human bone marrow cryopreservation.
    • The method maintains adequate viability of hematopoietic stem cells, as indicated by CFU-c efficiency.
    • This technique supports the potential for successful future clinical applications of cryopreserved bone marrow.