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Comparative Study between Three Methods of Stem Cell Separation from Cord Blood

Authors(4) :-Akram Deghady, Mona Wagdy Ayad , Yasser Orief, Mona Abo Bakr El-Shabassy

Background: Stem cells can be found in places like bone marrow and fat tissue, but the youngest, most flexible stem cells in the body come from the umbilical cord Several studies have shown the simplicity of umbilical cord blood collection however, stem cell separation method together with other factors such as unit size, maternal factors such as number of previous pregnancies age of the mother can affect the stem cells harvested from cord blood.
Aim of the work: Was to compare the outcome of three different methods for cord blood stem cells separation.
Material and methods: The study was carried out on 60 samples of umbilical cord blood collected from El Shatby Maternity Hospital, Faculty of Medicine, University of Alexandria.in this study we divided the samples into 3groups each of 20cord blood sample and used them to evaluate three separation methods: density gradient separation using Ficoll-Paque, an automated processing and harvesting system using AXP autoxpress thermogenesis, and magnetic bead separation method using MACS Columns and MACS Separators.
Results: There was a significant increase in the absolute number and the percentage of CD34 positive cells /total number of mononuclear cells   harvested by the magnetic beads method 190.0±87.6 cell/µl, (5.4±2.8 %) compared to those separated by AXP autoxpress Thermogenesis method 20.1±10.9 cell/µl (0.7±0.4%), and density gradient separation method 62.6±78.1 cell/µl. (1.8±1.9 % of total nucleated cells .2.7% to 12.2% (5.4±2.8) more efficient than the other two methods.
Conclusion: The magnetic bead separation method was proven to be the most efficient method for CD34 recovery among the 3 methods used.

Akram Deghady, Mona Wagdy Ayad , Yasser Orief, Mona Abo Bakr El-Shabassy
Stem Cells, Magnetic Beads, Cord Blood.
  1. Rosenthal J, Brown HL, Harris DT. Stem cell recovery following implementation of an automated cord blood processing system in a high volume laboratory. Biol Blood Marrow Transplant 2008;14(2):42s.
  2. Armson BA. Umbilical cord blood banking: implications for perinatal care providers. J Obstet Gynaecol Can 2005;27(3):263–90
  3. Barini R, Ferraz UC, Acácio GL, Machado IN. Does the time between collecting and processing umbilical cord blood samples affect the quality of the sample? Einstein. 2011;9(2):207–11
  4. Waters Ah. Platelet and granulocyte antigens and antibodies. In: Dacie JV, Lewis SM. Practical Haematology. 7th ed. London. Churchill Livingstone 1991;441-54
  5. Eunju O,Byung HL,Hyun-Young A, et al. Efficient nonadhesive ex vivo expansion of early endothelial progenitor cells derived from CD34+ human cord blood fraction for effective therapeutic vascularization. FASEB J 2011; 25: 159–169.
  6. Rubenstein P. Cord blood banking for clinical transplantation. Bone Marrow Transplantation 2009;44:635-642
  7. Etemadifar M, Dehghani L, Ganji H, Soleimani R, Talebi M, Eskandari N, Samani FS, Meamar R. Evaluation of the circulating CD34 + , CD309 + , and endothelial progenitor cells in patients with first attack of optic neuritis. Adv Biomed Res 2015,  4:151
  8. MiltenyiS , Schmitz J .In: Radbruch A (ed). Flowcytometry and cell sorting. Berlin Heidelberg New York. Springer. 1999; p 218
  9. Chalmers JJ, Zborowski M, Sun L, Moore L. Flow through, immunomagnetic cell separation. Biotechnol Prog. 1998;14(1): 141-148
  10. Pierzchalski A, Mittag A, Bocsi J, Tarnok A .An Innovative Cascade System for Simultaneous Separation of Multiple Cell Types. PLoS ONE 2013; 8(9): e74745
  11. Pafumi C, Bosco P, Cavallaro A, Farina M, Leonardi I, et al.  Two CD34+ stem cells from umbilical cord blood enrichment methods. Pediatr Hematol Oncol 2002; 19: 239–245
  12. Imai Y et al. Isolation and transplantation of highly purified autologous peripheral CD34 progenitor cells: purging efficacy, hematopoietic reconstitution in non-Hodgkin’s lymphoma (NHL). Bone Marrow Transplantation (2005) 35, 479–487
  13. Wynter de EA, Coutinho LH, Pei X ,et al. Comparison of purity and enrichment of CD34 cells from bone marrow, umbilical cord and peripheral blood (primed for apheresis) using five separation systems. Stem Cells 1995; 13: 524–532
  14. Kato K, Radbruch A. Isolation and characterization of CD34 hematopoietic stem cells from human peripheral blood by high-gradient magnetic cell sorting. Cytometry 1993; 14:384–392
  15. Querol S, Azqueta C, Garcia J. Effect of red blood cell content on progenitor function alter cryopreservation of cord blood buffy coat products. Bone Marrow Transplant 2002; 29 (S2): S202
  16. Lapierre V, Pellegrini N, Bardey I, et al. Cord blood volume reduction using an automated system (Sepax) vs. a semiautomated system (Optipress II) and a manual method (hydroxyethyl starch sedimentation) for routine cord blood banking: a comparative study. Cytotherapy 2007; 9: 165-9
  17. American Association of Blood Banks (AABB). Standards for Cellular Therapy Product Services, 5th ed.; AABB Press: Bethesda, MD, USA, 2012
  18. Boyum A.Isolation of leucocytes from human blood. A two-phase system for removal of red cells with methylcellulose as erythrocyte-aggregating agent. Scand J Clin Lab Invest Suppl. 1968; 97: 9-29.
  19. Kurnick JT, Ostberg L, Stegagno M, Kimura AK, Orn A,  Sjöberg O. A rapid method for the separation of functional lymphoid cell populations of human and animal origin on PVP-silica (Percoll) density gradients. Scand J Immunol. 1979; 10(6): 563-573.
  20. Warkentin PI, Hilden JM, Kersey JH, Ramsay NK, McCullough J. Transplantation of major ABO-incompatible bone marrow depleted of red cells by hydroxyethyl starch. Vox Sang. 1985; 48(2): 89-104.
  21. Smith RJ, Roath S. Bone marrow processing for transplantation using a Fenwal CS3000 Plus cell separator and a revised version of the Georgetown University procedure: comparison with a starch sedimentation method. Prog Clin Biol Res. 1994; 389: 705-710.
Publication Details
  Published in : Volume 2 | Issue 3 | May-June 2016
  Date of Publication : 2017-12-31
License:  This work is licensed under a Creative Commons Attribution 4.0 International License.
Page(s) : 85-90
Manuscript Number : IJSRST162323
Publisher : Technoscience Academy
PRINT ISSN : 2395-6011
ONLINE ISSN : 2395-602X
Cite This Article :
Akram Deghady, Mona Wagdy Ayad , Yasser Orief, Mona Abo Bakr El-Shabassy, "Comparative Study between Three Methods of Stem Cell Separation from Cord Blood", International Journal of Scientific Research in Science and Technology(IJSRST), Print ISSN : 2395-6011, Online ISSN : 2395-602X, Volume 2, Issue 3, pp.85-90, May-June-2016
URL : http://ijsrst.com/IJSRST162323