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Technical Aspects and Clinical Impact of Hematopoietic Progenitor Subset Quantification

^a The Department of Clinical Immunology and Transfusion Medicine, Aalborg Hospital;
^b The Department of Hematology, Herlev Hospital, University of Copenhagen, Copenhagen, Denmark

Key Words. CD34 • Analysis • Flow cytometry • Differentiation antigens • Cytokine receptors • Adherence molecules • Hematopoietic stem cell transplantation • Prediction of engraftment

John Baech, M.D., The Department of Clinical Immunology and Transfusion Medicine, Aalborg Hospital, Postbox 561, DK-9100 Aalborg, Denmark. Telephone: 45-99-32-11-11; Fax: 45-99-32-11-39; e-mail: john{at}dadlnet.dk

As high-dose therapy for malignancies is now being applied to newly diagnosed patients as adjuvant therapy, it has become a requirement that quality and safety assessment of hematopoietic stem cell grafts be evidence-based. This process has developed a new institution in medicine, the stem cell laboratory. In most cases this speciality has evolved from or within hematological research laboratories. However, the increased routine technologies applied in quality evaluation, ex vivo manipulation and safety assessment in stem cell handling naturally places this activity in transfusion medicine.

Multiparametric flow cytometry can identify progenitor subsets in normal human bone marrow and peripheral blood, and such subset quantification has been used retrospectively to predict three-lineage engraftment following high-dose therapy for malignancies. Published single center data have suggested an impact on clinical outcome, and a standardized technique for subset enumeration needs to be established before prospective multicenter trials can be initiated to document the prognostic value of such quality assessment in autografting.

Based on experiences of CD34 enumeration, which we consider to be the first step in quality assessment of hematopoietic stem cell grafts, this review discusses flow cytometry subset identification by lineage-specific differentiation markers, stromal-dependent adherence molecules, and regulatory growth factor receptors from a technical point of view. The aim of this review is:

• To recommend a simple method based on the experiences of the Nordic workshop III on subset identification;
  
• To present new molecular genetic-based methods for future use in quality assessment; and
  
• To propose new endpoints necessary for validation of the likely clinical impact of subsets in prospective trials.
  

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