Quantification of CD34+ cells: comparison of methods

Fritsch, Gerhard; Printz, Dieter; Stimpfl, Margit; Dworzak, Michael; Witt, Volker; Pötschger, Ulrike; Buchinger, P

doi:10.1046/j.1537-2995.1997.37897424398.x

Cited by 41 publications

(39 citation statements)

References 19 publications

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“…24 Also of importance are the nature of the CD34 mAb, the gating strategy 15,25,16 (particularly the use of an anti-CD45 mAb to identify leukocytes), and the number of counted events. 16,25,26 However, at least two studies in which variability in flow cytometry data was assessed, suggest that differences between distinct laboratories may be clinically insignificant.…”

Section: Discussionmentioning

confidence: 99%

CD34+ cell enumeration in peripheral blood and apheresis samples, using two laboratory diagnostic kits or an institutional protocol

Olivero

Alario

Ladaique

et al. 1999

Bone Marrow Transplant

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Summary:In order to prepare the substitution of a commercially available diagnostic kit, ProCOUNT (Becton Dickinson) or Stem-Kit (Coulter Immunotech), for our institutional protocol, we compared the three techniques for the numeration of CD34 ؉ progenitor cells in 50 peripheral blood and 51 apheresis samples, obtained from cancer patients or healthy donors. We show here that the three techniques yield results of the same order of magnitude. Although statistical analyses demonstrate significant differences between the three methods, these differences turned out to be clinically insignificant in most situations. Observed differences mostly affect samples with the highest content of CD34 ؉ cells, while the three assays provide equivalent results for values that are close to clinically relevant thresholds (20 ؋ 10 3 CD34 ؉ cells/ml in peripheral blood to start apheresis, and accumulated number above 3 ؋ 10 6 CD34 ؉ cells/kg to stop apheresis). This study also supports the view that institutional protocols can provide a highly reliable determination of CD34 ؉ cells counts and percentages. However, because institutional protocols often use research reagents and vary from institution to institution, the use of diagnostic kits may be prefered as one way to improve quality assurance in the practice of cell therapy.

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Section: Discussionmentioning

confidence: 99%

CD34+ cell enumeration in peripheral blood and apheresis samples, using two laboratory diagnostic kits or an institutional protocol

Olivero

Alario

Ladaique

et al. 1999

Bone Marrow Transplant

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show abstract

“…Cell preparation and immunological staining were done as described elsewhere. 7 A FACSCalibur (Becton Dickinson) equipped with the CELLQUEST and the PAINT-A-GATE PRO softwares (Becton Dickinson) was used for data acquisition and analysis. 7 …”

Section: Methodsmentioning

confidence: 99%

Resolution of early cytomegalovirus (CMV) infection after leukocyte transfusion therapy from a CMV seropositive donor

Witt

Fritsch

Peters

et al. 1998

Bone Marrow Transplant

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Summary:A 2 year and 8 month old CMV-negative boy suffering from stage III neuroblastoma underwent ABMT in first very good partial remission. He acquired early CMV infection on day +5, followed by consecutive graft failure and severe sepsis, and the clincial course deteriorated. Between days +16 and +21, he received seven leukocyte concentrates (LC) collected from a healthy, but CMV-IgG-seropositive relative stimulated with G-CSF (filgastrim, 5 g/kg/day). A median of 5.7 × 10 10 neutrophils/m 2 /day (range, 1.2-8.3) were transfused, corresponding to a T cell number of roughly 4 × 10 8 CD3 + cells/kg/day. After infusion of the LCs, PCR analysis became negative for CMV and the patient received his rescue bone marrow. One year after ABMT, he is in complete remission and in good clinical condition. Our results suggest that the T cells infused together with the irradiated leukocytes played a major role in eradicating the CMV infection in this patient.

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“…30 The Paint-A-Gate software (BD) was employed for data evaluation. FACS sorting of leucocyte subpopulations on the FACSVantage SE (BD) was performed after three-colour staining in a lyse-and-wash procedure.…”

Section: Facs Analysis and Cell Sortingmentioning

confidence: 99%

Lineage-specific chimaerism after stem cell transplantation in children following reduced intensity conditioning: potential predictive value of NK cell chimaerism for late graft rejection

et al. 2003

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Chimaerism of FACS-sorted leucocyte subsets (CD14 þ , CD15 þ , CD3À/56 þ , CD3 þ /4 þ , CD3 þ /8 þ , CD19 þ ) was monitored prospectively between days þ 14 and þ 100 in 39 children undergoing allogeneic stem cell transplantation with reduced intensity-conditioning regimens. Cell subsets exceeding 1% of nucleated cells were subject to cell sorting. Chimaerism was analysed by dual-colour FISH and/or by short tandem repeat-polymerase chain reaction. The chimaerism pattern on day þ 28 was evaluated with regard to its correlation with graft rejection. Of 39 patients, nine patients had donor chimaerism (DC) in all subsets. Mixed/recipient chimaerism (MC/RC) was detectable within T cells in 62%, within NK cells in 39% and within monocytes and granulocytes in 38% of the patients. The correlation of secondary graft rejection with the chimaerism pattern on day þ 28 revealed the strongest association between RC in NK-cells (Po0.0001), followed by T cells (P ¼ 0.001), and granulocytes and monocytes (P ¼ 0.034). Notably, patients with RC in T cells rejected their graft only if MC or RC was also present in the NK-cell subset. By contrast, none of the children with DC in NK cells experienced a graft rejection. These observations suggest that, in the presence of recipient T-cell chimaerism, the chimaerism status in NK-cells on day þ 28 might be able to identify patients at high risk for late graft rejection.

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Quantification of CD34+ cells: comparison of methods

Abstract: Neither density separation nor the whole-blood lysis procedure seems appropriate for optimal CD34+ quantification.

Cited by 41 publications

References 19 publications

CD34+ cell enumeration in peripheral blood and apheresis samples, using two laboratory diagnostic kits or an institutional protocol

CD34+ cell enumeration in peripheral blood and apheresis samples, using two laboratory diagnostic kits or an institutional protocol

Resolution of early cytomegalovirus (CMV) infection after leukocyte transfusion therapy from a CMV seropositive donor

Lineage-specific chimaerism after stem cell transplantation in children following reduced intensity conditioning: potential predictive value of NK cell chimaerism for late graft rejection

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