A theoretically optimized method for cord blood stem cell cryopreservation

Erik J. Woods, Jun Liu, Karen Pollok, Jennifer Hartwell, Franklin O. Smith, David A. Williams, Mervin Yoder, John K. Critser

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

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°). 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.

Original languageEnglish
Pages (from-to)341-350
Number of pages10
JournalJournal of Hematotherapy and Stem Cell Research
Volume12
Issue number3
DOIs
StatePublished - Jun 2003

Fingerprint

Cryopreservation
Fetal Blood
Blood Cells
Stem Cells
Dimethyl Sulfoxide
Permeability
Inbred NOD Mouse
SCID Mice
Temperature
Polychlorinated Biphenyls
Hematopoietic Stem Cells
B-Lymphocytes
Theoretical Models
Nitrogen
Water

ASJC Scopus subject areas

  • Hematology
  • Immunology

Cite this

A theoretically optimized method for cord blood stem cell cryopreservation. / Woods, Erik J.; Liu, Jun; Pollok, Karen; Hartwell, Jennifer; Smith, Franklin O.; Williams, David A.; Yoder, Mervin; Critser, John K.

In: Journal of Hematotherapy and Stem Cell Research, Vol. 12, No. 3, 06.2003, p. 341-350.

Research output: Contribution to journalArticle

Woods, Erik J. ; Liu, Jun ; Pollok, Karen ; Hartwell, Jennifer ; Smith, Franklin O. ; Williams, David A. ; Yoder, Mervin ; Critser, John K. / A theoretically optimized method for cord blood stem cell cryopreservation. In: Journal of Hematotherapy and Stem Cell Research. 2003 ; Vol. 12, No. 3. pp. 341-350.
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