Accessible Requires Authentication Published by De Gruyter June 1, 2005

The effect of processing and cryopreservation on nucleated umbilical cord blood cells

Lucy A. Bayer-Zwirello, Despina E. Hoffman, Lorrie A. Adams, Paul T. Wilder and Margaret T. Reece
From the journal


Recovery of nucleated cord blood cells after storage in liquid nitrogen was evaluated. Red cells were depleted using Ficoll-Paque™ or Puregene® red cell lyses. Freeze Medium contained 10% dimethylsulfoxide and 20% serum for cryoprotection. Recovery of the original cell population remaining serviceable for fluorescence activated cell sorting (FACS) was 12 ± 10% (average ± standard deviation), with a range of 1% to 55%. Viability measured by FACS analysis after freezing was significantly lower than that of the same specimens prior to freezing, 62 ± 20% compared to 91 ± 11% (p < 0.001). Percentage CD45+34+ cells were the same for fresh and frozen cells. Gestational age at which specimens were collected had no effect on the percent cells carrying the CD45+34+ markers. We conclude that better cryoprotective supplements are needed to insure consistent high recovery of viable nucleated umbilical cord blood cells after preservation in liquid nitrogen.


Corresponding author: Division of Maternal-Fetal Medicine, Department of Obstetrics and Gynecology, St. Elizabeth’s Medical Center, 736 Cambridge Street, Boston, MA 02135/U.S.A. E-mail:


1 Broxmeyer HE, EF Srour, G Hangoc, S Cooper, SA Anderson, DM Bodine: High-efficiency recovery of functional hematopoietic progenitor and stem cells from human cord blood cryopreserved for 15 years. Proc Natl Acad Sci USA100 (2003) 645 Search in Google Scholar

2 Donaldson C, WJ Armitage, PA Denning-Kendall, AJ Nicol, BA Bradley, JM Hows: Optimal cryopreservation of human umbilical cord blood. Bone Marrow Transplant20 (1997) 89 Search in Google Scholar

3 Forte L, G Migliaccio, M Sanchez, AR Migliaccio, AM Passarelli, S Amadori: Effects of cell banking manipulations on ex vivo amplification of umbilical cord blood. Am Ist Super Sanita36 (2000) 333 Search in Google Scholar

4 Galmes A, J Besalduch, J Bargay, A Novo, M Morey, JM Guerra, MA Duran: Long-term storage at –80°C of hematopoietic progenitor cells with 5-percent dimethyl sulfoxide as the sole cryoprotectant. Transfusion39 (1999) 70 Search in Google Scholar

5 Haneline LS, KP Marshall, DW Clapp: The highest concentration of primitive hematopoietic progenitor cells in cord blood is found in extremely premature infants. Pediatr Res39 (1996) 820 Search in Google Scholar

6 Keeney M, I Chin-Yee, K Weir, J Popma, R Nayar, DR Sutherland: Single platform flow cytometric absolute CD34+ cell counts based on the ISHAGE guidelines. Cytometry34 (1998) 61 Search in Google Scholar

7 Kobylka P, P Ivanyi, BS Breur-Vriesendorp: Preservation of immunological and colony-forming capacities of long-term (15 years) cryopreserved cord blood cells. Transplantation65 (1998) 1275 Search in Google Scholar

8 Lim F, J Beckhoven, A Brand, J Kluin-Nelemans, R Willemze, H Kanhai, J Falkenburg: The number of nucleated cells reflects the hematopoietic content of umbilical cord blood for transplantation. Bone Marrow Transplant24 (1999) 965 Search in Google Scholar

9 Rubinstein P, C Carrier, A Scaradavou, J Kurtzberg, J Adamson, AR Migliaccio, RL Berkowitz, M Cabbad, NL Dobrila, PE Taylor, RE Rosenfield, CE Stevens: Outcomes among 562 recipients of placental-blood transplants from unrelated donors. N Engl J Med339 (1998) 1565 Search in Google Scholar

10 Surbek DV, W Holzgreve, W Jansen, D Heim, H Garritsen, C Nissen, A Wodnar-Filipowicz: Quantitative immunophenotypic characterization, cryopreservation, and enrichment of second and third trimester human fetal cord blood hematopoietic stem cells (progenitor cells). Am J Obstet Gynecol179 (1998) 1228 Search in Google Scholar

Published Online: 2005-06-01
Published in Print: 2004-09-01

© Walter de Gruyter