Document Detail

Damage and protection of UC blood cells during cryopreservation.
MedLine Citation:
PMID:  11953017     Owner:  NLM     Status:  MEDLINE    
BACKGROUND: Current procedures for the cryopreservation of umbilical cord blood (UCB) progenitor cells, which are based on techniques used for BM, have had varying degrees of success (survival 9-118%). Improving the effectiveness of UCB cell therapies demands a more comprehensive understanding of freezing injury during cryopreservation. METHODS: Leukocyte concentrates from UCB, with or without 10% DMSO were cooled at 1 degrees C/min to different subzero temperatures (-5 to -50 degrees C), then either thawed directly (thaw) or plunged into liquid nitrogen before thawing (plunge). Single-platform flow cytometry with 7-amino-actinomycin D was used to directly quantify survival of CD34(+) cells. Fluorescent microscopy was used to examine plasma membrane integrity of nucleated cells. RESULTS: Without DMSO, recovery of nucleated cells was approximately 80% for both thaw and plunge. Survival was 9%, indicating damage to the plasma membrane. With 10% DMSO, nucleated cell recovery was also approximately 80%, indicating that DMSO does not improve recovery of nucleated cells. Survival, however, was much higher with DMSO, > 60% for nucleated cells thawed directly, and 30-55% for cells thawed from plunge, demonstrating cryoprotection conferred by DMSO. With DMSO, survival of CD34(+) cells was higher than that of nucleated cells, indicating that CD34(+) cells with 10% DMSO are more tolerant to cryopreservation than the total nucleated cell population. DISCUSSION: This study provides the necessary data on the low temperature response of UCB progenitor cells that are critical for the development of standards for the cryopreservation of UCB.
H Yang; J P Acker; J Hannon; H Miszta-Lane; J J Akabutu; L E McGann
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Cytotherapy     Volume:  3     ISSN:  1465-3249     ISO Abbreviation:  Cytotherapy     Publication Date:  2001  
Date Detail:
Created Date:  2002-04-15     Completed Date:  2003-01-24     Revised Date:  2008-04-24    
Medline Journal Info:
Nlm Unique ID:  100895309     Medline TA:  Cytotherapy     Country:  England    
Other Details:
Languages:  eng     Pagination:  377-86     Citation Subset:  IM    
Alberta Cord Blood Bank, Canadian Blood Services, Edmonton, Alberta, Canada.
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MeSH Terms
Antigens, CD34 / analysis,  immunology
Cell Survival / drug effects
Cryopreservation / methods*
Dimethyl Sulfoxide / pharmacology
Fetal Blood / cytology*,  drug effects,  metabolism
Flow Cytometry / methods
Hematopoietic Stem Cells / cytology,  drug effects,  metabolism,  pathology
Leukocytes / cytology,  drug effects
Reg. No./Substance:
0/Antigens, CD34; 67-68-5/Dimethyl Sulfoxide

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