A Theoretically Optimized Method for Cord Blood Stem Cell Cryopreservation

低温保存 二甲基亚砜 干细胞 化学 低温生物学 脐带血 男科 造血 低温保护剂 生物 医学 脐带 免疫学 细胞生物学 胚胎 有机化学
作者
Erik J. Woods,Jun Liu,Karen E. Pollok,Jennifer R. Hartwell,Franklin O. Smith,David A. Williams,Mervin C. Yöder,John K. Critser
出处
期刊:Journal of Hematotherapy & Stem Cell Research [Mary Ann Liebert]
卷期号:12 (3): 341-350 被引量:29
标识
DOI:10.1089/152581603322023070
摘要

The objective of this study was to develop an optimal cryopreservation method for human umbilical cord blood hematopoietic progenitor cells as evidenced by improved retention of in vivo engraftment ability and multilineage differentiation. An extended understanding of the osmometric/permeability characteristics of cord blood stem cells was accomplished by measuring permeability of the cryoprotectant dimethyl sulfoxide (DMSO) at below-ambient temperatures (10° and 3°C). These data were combined with previously published osmotic and permeability data and the water–NaCl–DMSO phase diagram in conjunction with a mathematical model to determine an optimal initial DMSO concentration, cooling rate, and liquid nitrogen plunging temperature. Cells cryopreserved with the theoretically optimized procedure were then compared with cells frozen using standard methods for the ability to engraft in irradiated NOD/SCID mice. The optimal procedure was determined to include a 0.7 molal (~5%) DMSO concentration at a cooling rate of 4°C/min, and a plunging temperature of –44°C. The optimized protocol resulted in significantly higher engraftment of human CD45+ cells (17.2 ± 1.6% vs. 8.4 ± 1.6%), CD19+ B lymphocytes (11.3 ± 1.2% vs. 5.8 ± 1.2%), and CD34+ cells (1.9 ± 0.09% vs. 0.6 ± 0.09%) compared to cells frozen using a standard method. Engraftment of CD33+ cells was not significantly different (4.0 ± 0.3 vs. 3.2 ± 0.6, respectively). This study demonstrated that the use of a theoretically determined optimal cryopreservation method is superior to standard methods for maintaining UCB PCBs with multilineage repopulation potential in NOD/SCID mice.
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