Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

Cell Procurement

Factors affecting volume reduction and red blood cell depletion of bone marrow on the COBE Spectra cell separator before haematopoietic stem cell transplantation

Abstract

The COBE Spectra™ is used to volume/red blood cell (RBC) deplete BM before transplantation or cryopreservation. We have audited our results to identify the effect of transit time, refrigerated storage, age and cellular composition on mononuclear cells (MNC) and CD34+ cell recoveries, volume/RBC depletion and neutrophil engraftment. In total, 88 consecutive collections from autologous (n=25) and allogeneic (n=63) donors were included. The mean collection volume was 1250±398 ml with RBC content of 341±113 ml. The MNC and CD34+ cell recoveries were 83.3±18.5 and 88.1±18.9%, respectively, volume depletion was 88.2±4.4% and RBC depletion 98.3±1.8%. Neutrophil engraftment was achieved in 20.1±6.4 days. Factors affecting MNC and CD34+ cell recoveries were transit time (P=0.0060), overall age (P<0.0210) and MNC/CD34+ cell concentrations (P<0.0313). The presence of crenated RBC also reduced CD34+ cell recovery (P=0.0028). Refrigerated storage did not adversely affect cell recovery (P>0.8161) or neutrophil engraftment (P=0.8959). This study demonstrates that time in transit, overall age, MNC and CD34+ cell concentrations and RBC condition were important factors affecting processing. RBCs show artefacts soon after collection at ambient temperatures and these may interfere with the separation and collection of MNC/CD34+ cells. Refrigeration at 4–6°C during transit and storage may reduce formation of RBC artefacts and maximize MNC and CD34+ cell recoveries.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2

Similar content being viewed by others

References

  1. Klumpp TR . Immunohematologic complications of bone marrow transplantation. Bone Marrow Transplantation 1991; 8: 159–170.

    CAS  PubMed  Google Scholar 

  2. Korbling M, Huh YO, Durett A, Mirza N, Miller P, Engel H et al. Allogeneic blood stem cell transplantation: peripheralization and yield of donor-derived primitive hematopoietic progenitor cells (CD34+ Thy-1dim) and lymphoid subsets, and possible predictors of engraftment and graft-versus-host disease. Blood 1995; 86: 2842–2848.

    CAS  Google Scholar 

  3. Bensinger WI, Longin K, Appelbaum F, Rowley S, Weaver C, Lilleby K et al. Peripheral blood stem cells (PBSCs) collected after recombinant granulocyte colony stimulating factor (rhG-CSF): an analysis of factors correlating with the tempo of engraftment after transplantation. Br J Haematol 1995; 87: 825–831.

    Article  Google Scholar 

  4. Baker PK, Rhodes EG, Duguid JK . Continuous flow cell separator use for bone marrow processing. Transfus Sci 1991; 12: 183–187.

    Article  CAS  Google Scholar 

  5. Koristek Z, Mayer J . Bone marrow processing for transplantation using the COBE Spectra cell separator. J Hematother Stem Cell Res 1999; 8: 443–448.

    Article  CAS  Google Scholar 

  6. Cleaver SA, Warren P, Kern M, Hurley CK, Raffoux C, Keller J et al. Donor work-up and transport of bone marrow – recommendations and requirements for a standardized practice throughout the world from donor Registries and Quality Assurance Working Groups of the World Marrow Donor Association (WMDA). Bone Marrow Transplant 1997; 20: 621–629.

    Article  CAS  Google Scholar 

  7. Keeney M, Chin-Yee I, Weir K, Popma J, Nayar R, Sutherland DR . Single platform flow cytometric absolute CD34+ cell counts based on the ISHAGE guidelines. International Society of Hematotherapy and Graft Engineering. Cytometry 1998; 34: 61–70.

    Article  CAS  Google Scholar 

  8. Hollyman D, Sidders C, Lankester A, Watt S, Pamphilon D, Guttridge MG . A novel lyse-no-wash 4-colour flow cytometry assay with an internal positive control for the simultaneous enumeration of CD34+ and CD3+ cells prior to allogeneic transplantation. (ISCT abstract 230 http://www.celltherapy.org/Abstracts ).

  9. Pamphilon DH . Transfusion policy. In: Apperley J, Carreras E, Gluckman E, Gratwohl A, Masszi T (eds). The EBMT Handbook – Haematopoietic Stem Cell Transplantation. Forum Service Editore: Genoa, 2004, pp 107–117.

    Google Scholar 

  10. Lewis SM . Collection and handling of blood. In: Lewis SM, Bain BJ, Bates I (eds). Dacie and Lewis Practical Haematology. Churchill Livingstone: London, 2001, pp 1–8.

    Google Scholar 

  11. Brittin GM, Brecher G, Johnson CA, Elashoff RM . Stability of blood in commonly used anticoagulants. Am J Clin Pathol 1969; 52: 690–694.

    Article  CAS  Google Scholar 

  12. Lewis SM . Reference ranges and normal values. In: Lewis SM, Bain BJ, Bates I (eds). Dacie and Lewis Practical Haematology. Churchill Livingstone: London, 2001, pp 9–18.

    Google Scholar 

  13. Lawrence AC, Bevington JM, Young M . Storage of blood and the mean corpuscular volume. J Clin Pathol 1975; 28: 345–349.

    Article  CAS  Google Scholar 

  14. Lioznov MV, Freiberger P, Kröger N, Zander AR, Fehse B . Aldehyde dehydrogenase activity as a marker for the quality of hematopoietic stem cell transplants. Bone Marrow Transplant 2005; 35: 909–914.

    Article  CAS  Google Scholar 

  15. Stainsby D, Cohen H, Jones H, Boncinelli A, Knowles S, Birchall J et al. Serious Hazards of Transfusion Annual Report. 2004. http://www.shot-uk.org.

Download references

Acknowledgements

This work is supported by the NHS Blood and Transplant Authority and the Department of Health, UK.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to M G Guttridge.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Guttridge, M., Sidders, C., Booth-Davey, E. et al. Factors affecting volume reduction and red blood cell depletion of bone marrow on the COBE Spectra cell separator before haematopoietic stem cell transplantation. Bone Marrow Transplant 38, 175–181 (2006). https://doi.org/10.1038/sj.bmt.1705420

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.bmt.1705420

Keywords

This article is cited by

Search

Quick links