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Trehalose ameliorates the cryopreservation of cord blood in a preclinical system and increases the recovery of CFUs, long-term culture-initiating cells, and nonobese diabetic-SCID repopulating cells

Authors

  • Xiao Bing Zhang,

    1. From the Department of Paediatrics and the Department of Anatomical and Cellular Pathology, Prince of Wales Hospital, The Chinese University of Hong Kong, Hong Kong.
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  • Karen Li,

    1. From the Department of Paediatrics and the Department of Anatomical and Cellular Pathology, Prince of Wales Hospital, The Chinese University of Hong Kong, Hong Kong.
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  • Kwai Heung Yau,

    1. From the Department of Paediatrics and the Department of Anatomical and Cellular Pathology, Prince of Wales Hospital, The Chinese University of Hong Kong, Hong Kong.
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  • Kam Sze Tsang,

    1. From the Department of Paediatrics and the Department of Anatomical and Cellular Pathology, Prince of Wales Hospital, The Chinese University of Hong Kong, Hong Kong.
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  • Tai Fai Fok,

    1. From the Department of Paediatrics and the Department of Anatomical and Cellular Pathology, Prince of Wales Hospital, The Chinese University of Hong Kong, Hong Kong.
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  • Chi Kong Li,

    1. From the Department of Paediatrics and the Department of Anatomical and Cellular Pathology, Prince of Wales Hospital, The Chinese University of Hong Kong, Hong Kong.
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  • Shuk Man Lee,

    1. From the Department of Paediatrics and the Department of Anatomical and Cellular Pathology, Prince of Wales Hospital, The Chinese University of Hong Kong, Hong Kong.
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  • Patrick Man Pan Yuen

    1. From the Department of Paediatrics and the Department of Anatomical and Cellular Pathology, Prince of Wales Hospital, The Chinese University of Hong Kong, Hong Kong.
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  • ABBREVIATIONS:

    CB = cord blood; CFU-GEMM = CFU-granulocyte erythroid macrophage megakaryocytes; CFU-MK = CFU-megakaryocyte; IMDM = Iscove's modified Dulbecco's medium; LTC-IC = long-term culture-initiating cells; NC = nucleated cell; NOD-SCID = nonobese diabetic-SCID; PB = peripheral blood.

  • This study was supported by the Industrial Support Fund #AF/203/98, Department of Industry, Hong Kong Government Special Administrative Region.

Address reprint requests to: Karen Li, PhD, Depart- ment of Paediatrics, 6/F, Clinical Sciences Building, Prince of Wales Hospital, Shatin, N. T., Hong Kong. e-mail: lipang@cuhk.edu.hk .

Abstract

BACKGROUND : The cryopreservation of HPCs in DMSO has been practiced by cord blood (CB) banks worldwide. Inevitably, some detriment to biologic function occurs as the result of freezing injuries and DMSO toxicity. Trehalose, a disaccharide, is a natural cryoprotectant in organisms capable of surviving extreme dehydration and cold. The objective of this study was to establish the cryopreservation of CB under preclinical conditions using trehalose as a supplement to DMSO.

STUDY DESIGN AND METHODS : In a preclinical protocol, the effects of 5-percent trehalose with 10-percent DMSO or 5-percent DMSO on the cryopreservation of CB MNCs or nucleated cells (NCs) were further evaluated. The read-out system consisted of a panel of HPCs: early progenitors (CFU-GEMM, long-term culture-initiating cells [LTC-IC]) and committed progenitors (CFU-GM, CFU/BFU-E, CFU-megakaryocyte [CFU-MK]). The homing and engraftment capacity of these cells were assessed in nonobese diabetic (NOD)-SCID mice.

RESULTS : Trehalose increased the recoveries of CFU-GM, CFU/BFU-E, CFU-GEMM, and LTC-IC by over 7.25 percent (mean), 11.9 percent, 19.2 percent, and 12.9 percent, respectively, when compared with those in paired CB samples cryopreserved in 10-percent DMSO. Freezing and thawing reduced the yields of CFU-MK by 35.5 percent (mean) and 28.4 percent in MNC and NC samples, respectively, and the inclusion of 5-percent trehalose significantly retrieved these progenitor cells to over 90 percent of fresh samples. The improved recovery of functional HPLs was reflected by their multilineage engraftment in NOD-SCID mice.

CONCLUSION : Trehalose at 5 percent significantly ameliorates the cryopreservation of CB progenitor cells at a preclinical protocol. The increased recoveries of these cells might potentially improve the engraftment outcomes of CB transplants.

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