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Some early differentiation markers detected in cytoplasm of pre-B cells by flow cytometry.
Neoplasma. 1996; 43(6):373-9.N

Abstract

Immunofluorescent staining of cytoplasmic IgM (heavy chains) and CD24 as well as their simultaneous staining with surface B cell markers was used to study immunophenotype changes in B cell differentiation. Human hematopoietic B cell lines P3HR1 and RAJI were used. We found that IgM and CD24 cell markers while absent on cell membrane could be detected in their cytoplasm (c). The presence of cIgM in cell lines RAJI, P3HRI indicates their early pre-B differentiation stage. The presence of cCD24 simultaneously with mCD22 and cIgM is the evidence that hematopoietic cell lines or leukemias may not accurately reflect normal differentiation pathway. Combinations of cIgM, cCD24 with surface B cell markers CD10, CD19 on these cell lines can be considered as leukemia associated phenotypes. Some of them were shown in bone marrow and peripheral blood of pre-B ALL and B-CLL patients and can be used for the detection of minimal residual disease. Different fixation/permeabilization methods were tested in order to choose the optimal one for simple detection of cytoplasmic markers or their simultaneous detection with surface markers by flow cytometry. They included "one-component-methods" (methanol-M, saponin-S), methods combining these components with paraformaldehyde (P+M, P+S) or buffered formaldehyde acetone (BFA). The choice depended on individual marker detected. General parameters like the proportion of debris, cell aggregation, cell loss and the changes of scatter parameters FSC and SSC were taken into consideration. The priorities of combined methods P+S, P+M1 and BFA over one-component methods are demonstrated.

Authors+Show Affiliations

Cancer Research Institute, Slovak Academy of Sciences, Bratislava, Slovakia.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

8996561

Citation

Koníková, E, et al. "Some Early Differentiation Markers Detected in Cytoplasm of pre-B Cells By Flow Cytometry." Neoplasma, vol. 43, no. 6, 1996, pp. 373-9.
Koníková E, Glasová M, Kusenda J, et al. Some early differentiation markers detected in cytoplasm of pre-B cells by flow cytometry. Neoplasma. 1996;43(6):373-9.
Koníková, E., Glasová, M., Kusenda, J., & Babusíková, O. (1996). Some early differentiation markers detected in cytoplasm of pre-B cells by flow cytometry. Neoplasma, 43(6), 373-9.
Koníková E, et al. Some Early Differentiation Markers Detected in Cytoplasm of pre-B Cells By Flow Cytometry. Neoplasma. 1996;43(6):373-9. PubMed PMID: 8996561.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Some early differentiation markers detected in cytoplasm of pre-B cells by flow cytometry. AU - Koníková,E, AU - Glasová,M, AU - Kusenda,J, AU - Babusíková,O, PY - 1996/1/1/pubmed PY - 1996/1/1/medline PY - 1996/1/1/entrez SP - 373 EP - 9 JF - Neoplasma JO - Neoplasma VL - 43 IS - 6 N2 - Immunofluorescent staining of cytoplasmic IgM (heavy chains) and CD24 as well as their simultaneous staining with surface B cell markers was used to study immunophenotype changes in B cell differentiation. Human hematopoietic B cell lines P3HR1 and RAJI were used. We found that IgM and CD24 cell markers while absent on cell membrane could be detected in their cytoplasm (c). The presence of cIgM in cell lines RAJI, P3HRI indicates their early pre-B differentiation stage. The presence of cCD24 simultaneously with mCD22 and cIgM is the evidence that hematopoietic cell lines or leukemias may not accurately reflect normal differentiation pathway. Combinations of cIgM, cCD24 with surface B cell markers CD10, CD19 on these cell lines can be considered as leukemia associated phenotypes. Some of them were shown in bone marrow and peripheral blood of pre-B ALL and B-CLL patients and can be used for the detection of minimal residual disease. Different fixation/permeabilization methods were tested in order to choose the optimal one for simple detection of cytoplasmic markers or their simultaneous detection with surface markers by flow cytometry. They included "one-component-methods" (methanol-M, saponin-S), methods combining these components with paraformaldehyde (P+M, P+S) or buffered formaldehyde acetone (BFA). The choice depended on individual marker detected. General parameters like the proportion of debris, cell aggregation, cell loss and the changes of scatter parameters FSC and SSC were taken into consideration. The priorities of combined methods P+S, P+M1 and BFA over one-component methods are demonstrated. SN - 0028-2685 UR - https://www.unboundmedicine.com/medline/citation/8996561/Some_early_differentiation_markers_detected_in_cytoplasm_of_pre_B_cells_by_flow_cytometry_ DB - PRIME DP - Unbound Medicine ER -