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.
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.


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.


CONCLUSIONS
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.
Zhang
Xiao Bing
XB
Department of Paediatrics, Prince of Wales Hospital, The Chinese University of Hong Kong, Hong Kong.
Li
Karen
K
Yau
Kwai Heung
KH
Tsang
Kam Sze
KS
Fok
Tai Fai
TF
Li
Chi Kong
CK
Lee
Shuk Man
SM
Yuen
Patrick Man Pan
PM
D000818:Animals; D001793:Blood Preservation / Q000379:methods*; D019070:Cell Lineage; D003114:Colony-Forming Units Assay; D036101:Cord Blood Stem Cell Transplantation; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology*; D005312:Fetal Blood / Q000166:cytology* / Q000187:drug effects; D006085:Graft Survival; D006410:Hematopoiesis; D006412:Hematopoietic Stem Cells / Q000166:cytology; D006801:Humans; D017493:Leukocyte Common Antigens; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D014183:Transplantation, Heterologous; D014199:Trehalose / Q000494:pharmacology*
D003451:Cryoprotective Agents; D014199:Trehalose; D017493:Leukocyte Common Antigens