Immunomagnetic Positive Selection and Colony Culture of CD34<sup>+</sup> Cells from Blood

Law, Ping; Ishizawa, Lori; Ven, Carmella van de; Burgess, Julie; Hardwick, Alan; Plunkett, Michael; Gee, Adrian P.; Cairo, Mitchell S.

doi:10.1089/scd.1.1993.2.247

Cited by 18 publications

(13 citation statements)

References 3 publications

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“…CD34+ cells were purified by an immunomagnetic method (Isolex system, Baxter, Santa Ana, Calif., USA), as described elsewhere [9]. This method does not affect the surface phenotype or the potential of these CD34+ cells for engraftment [10].…”

Section: Methodsmentioning

confidence: 99%

Determination of Hematopoietic Stem Cells in Peripheral Blood by an Automated Hematology Analyzer (SE-9000)

Takekawa¹,

Yamane²,

Hino³

et al. 1998

Acta Haematol

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We evaluated the usefulness of an automated hematology analyzer (SE-9000) for the identification and counting of peripheral blood stem cells (PBSCs). The samples tested were from 14 patients with hematological malignancies. Peripheral blood samples were collected from the subjects before and after a course of chemotherapy. From the leukapheresis sample, CD34+ cells, assumed to be hematopoietic stem cells, were obtained with an immunomagnetic cell separator. The CD34+ cells obtained accumulated in the gate corresponding to low recurrent frequencies of the automated hematology analyzer. This gate shows results of the ‘immature information’ (IMI) channel. Software for detection of only the cells that accumulated in this gate was therefore developed. With this trial program, the regression coefficient between the percentage of leukocytes from the blood samples that were CD34+ and the percentage of such leukocytes that appeared on the IMI channel was 0.79. With this analyzer, the number of PBSC could be counted in about 80 s. The identification and counting of cells picked up by the IMI channel should be clinically useful for the monitoring of changes in PBSC after chemotherapy for mobilization.

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Section: Methodsmentioning

confidence: 99%

Determination of Hematopoietic Stem Cells in Peripheral Blood by an Automated Hematology Analyzer (SE-9000)

Takekawa¹,

Yamane²,

Hino³

et al. 1998

Acta Haematol

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Section: Introductionmentioning

confidence: 99%

“…Immunomagnetic systems have been used for positive selection in bone marrow in preclinical and clinical settings, with satisfactory results (2,7,8). However, when used with mobilized peripheral blood, these systems have achieved lower final purities than obtained with bone marrow or cord blood and, therefore, a lower tumor depletion efficiency (2,(7)(8)(9). The reasons for this are unclear, but different groups have proposed an additional predepletion step to try to improve the tumor depletion ability of positive selections from mobilized peripheral blood (9)(10)(11).…”

Section: Introductionmentioning

confidence: 99%

CD34+ Cell Positive Selection from Mobilized Peripheral Blood by an Indirect Immunomagnetic Method: Effect of the Type of Mobilization and Assessment of Tumor Depletion Ability

Cancelas¹,

Querol²,

Canals³

et al. 1995

Journal of Hematotherapy

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Mobilized peripheral blood has been shown to be a suitable source of hematopoietic progenitor cells for autologous transplantation in oncologic patients. However, tumor cell contamination can potentially occur, although to a lesser extent than in the bone marrow. CD34+ cell positive selection has been developed as a system for the ex vivo purging of remaining tumor cells when used in mobilized peripheral blood. This method has shown a lower purification potential than that obtained with bone marrow or cord blood. The reason for this is not clear, but different groups have tried to improve the purity and yield of the positive selection on mobilized peripheral blood by predepletion of nonlymphoid cell populations, since they can induce nonspecific binding. The present study was designed to test an indirect immunomagnetic CD34+ cell selection method to make it reproducible, feasible, and effective for purging mobilized peripheral blood. Twenty-nine samples from mobilized peripheral blood were tested. The median starting CD34+ percentage was 0.8 (0.3%-4.2%), and the median final purity was 87% (32.7%-99.7%), with a median yield of 44.8% (15%-83.5%). The highest purity was reproducibly achieved when the starting percentage of CD34+ cells was higher than 0.65% (median purity 93.7, range 81%-99.7%, CV 6%) on samples obtained from patients primed with chemotherapy alone or chemotherapy plus recombinant human granulocyte-colony stimulating factor. No relation was found between the content of contaminating nucleated cells and the final CD34+ cell purity. This method showed a depletion capacity, assessed by PCR on samples contaminated with K562 leukemic cells, of about 3 logs. These results indicate that this indirect immunomagnetic method can produce high purity CD34+ cell fractions from mobilized peripheral blood with a high efficiency of tumor depletion.

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“…17,18 In brief, cells were labeled with the murine anti-CD34 MoAb, 9C5 (Immunotherapy Division, Baxter Healthcare). Then, the cells attached to magnetic Patients (Table 1) beads coated with goat anti-mouse IgG antibody were colInformed consent was obtained from the patients' legal lected by magnets.…”

Section: Methodsmentioning

confidence: 99%

Allogeneic peripheral stem cell transplantation using positively selected CD34+ cells from HLA-mismatched donors

Matsuda

Hara

Osugi

et al. 1998

Bone Marrow Transplant

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Summary:and a preserved host immune system. Data on the effect of HLA mismatching on the incidence of graft failure showed We examined five children who underwent allogeneic an overall increase in the incidence in the mismatched peripheral stem cell transplantation (PSCT) using posigroup compared to HLA-identical sibling bone marrow tively selected CD34 ؉ cells from three or two loci-mistransplantation (BMT) and the frequency of graft failure matched donors. CD34 ؉ cells mobilized from peripheral increased as donor/recipient HLA disparity increased. 3,4 blood were separated by immunomagnetic beads.While the host immune system is defeated by T cells con-CD34 ؉ cells at 2.2-6.2 ؋ 10 6 /kg were transplanted into tained in the grafts, in the case of SCT with T cell-depleted three patients with refractory leukemia, a patient with grafts, immune competent cells in recipients are preserved. relapsed medulloblastoma and a patient with Fanconi's In the mouse model, the immunologic rejection of T cellanemia following a conditioning regimen which depleted histoincompatible BMT can be overcome by included irradiation, alkylating agents and antithymoincreasing the doses of irradiation or by adding selective cyte globulin treatment. The number of infused CD3 ؉ measures with minimal toxicity, such as splenic irradiation cells included in grafts was 2.3-22.7 ؋ 10 4 /kg. Four or in vivo treatment with anti-T cell monoclonal antibodies, patients achieved engraftment and hematopoietic reconto the conditioning regimen. 5-7 However, previous results stitution (Ͼ5 ؋ 10 8 /l of neutrophils on day 10 or 11).of clinical trials using these procedures have not been satisGraft rejection was observed in the patient with Fanfactory. 8,9 Another means of overcoming graft failure is to coni's anemia, but a rapid engraftment was obtained increase the number of stem cells in the graft. This has been after second PSCT. Although no prophylactic agents found to be the case in studies investigating the relationship other than ATG (included in the conditioning regimen) between transfused cell doses and the incidence of rejecwere used, greater than grade I acute GVHD was not tion. 10 Recently, successful engraftment using positively observed, but limited chronic GVHD was observed in selected allogeneic CD34 + bone marrow cells from two or two patients. The two patients with leukemia relapsed three loci-mismatched donors was reported. This procedure on days 103 and 210, respectively, and the patient with enables a larger number of transplantable stem cells. 11,12 medulloblastoma died of disease on day 159. The patientThe administration of recombinant human granulocyte with Fanconi's anemia died of fungal infection. CMV colony-stimulating factor (rhG-CSF) can mobilize a sufand HHV-6 diseases developed in four and two patients, ficient number of peripheral stem cells (PSCs) to permit respectively. Thus, although SCT using positively selecthe collection of a transplant inoculum. Infusion of these ted peripheral CD34 ؉ cells may be an alternative cytokine-...

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Immunomagnetic Positive Selection and Colony Culture of CD34⁺ Cells from Blood

Cited by 18 publications

References 3 publications

Determination of Hematopoietic Stem Cells in Peripheral Blood by an Automated Hematology Analyzer (SE-9000)

Determination of Hematopoietic Stem Cells in Peripheral Blood by an Automated Hematology Analyzer (SE-9000)

CD34+ Cell Positive Selection from Mobilized Peripheral Blood by an Indirect Immunomagnetic Method: Effect of the Type of Mobilization and Assessment of Tumor Depletion Ability

Allogeneic peripheral stem cell transplantation using positively selected CD34+ cells from HLA-mismatched donors

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Immunomagnetic Positive Selection and Colony Culture of CD34+ Cells from Blood

Cited by 18 publications

References 3 publications

Determination of Hematopoietic Stem Cells in Peripheral Blood by an Automated Hematology Analyzer (SE-9000)

Determination of Hematopoietic Stem Cells in Peripheral Blood by an Automated Hematology Analyzer (SE-9000)

CD34+ Cell Positive Selection from Mobilized Peripheral Blood by an Indirect Immunomagnetic Method: Effect of the Type of Mobilization and Assessment of Tumor Depletion Ability

Allogeneic peripheral stem cell transplantation using positively selected CD34+ cells from HLA-mismatched donors

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Immunomagnetic Positive Selection and Colony Culture of CD34⁺ Cells from Blood