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Assessment of cell proliferation in normal and pathological bone marrow biopsies: a study using double sequential
W Pellegrini1, F Facchetti, D Marocolo
1Department of Pathology, University of Brescia, Italy.
Insights
Proliferative activity in bone marrow cells, assessed using Ki-67 staining, shows similar erythroid and myeloid precursor activity across normal and myeloproliferative disorders. Increased megakaryocyte proliferation aids diagnosis, while myeloma shows low activity.
Area of Science:
- Hematology
- Immunohistochemistry
- Oncology
Background:
- Assessing bone marrow cell proliferation is crucial for diagnosing hematological disorders.
- Ki-67 antigen is a widely used marker for cell proliferation.
- Sequential double immunostaining allows simultaneous evaluation of proliferation and cell lineage.
Purpose of the Study:
- To evaluate the proliferative activity of hematopoietic and plasma cells in normal and neoplastic bone marrow conditions.
- To determine the utility of Ki-67 expression in differentiating various myeloproliferative disorders and myeloma.
- To assess the diagnostic value of MIB-1 and lineage-specific markers in routine bone marrow biopsies.
Main Methods:
- Analyzed 58 bone marrow biopsies (11 normal, 47 neoplastic) using sequential double immunostaining.
- Utilized MIB-1 antibody for Ki-67 antigen and antibodies for glycophorin-C, myeloperoxidase, factor VIII-related antigen, and immunoglobulin light chains.
- Quantified Ki-67 positive cells in erythroid precursors, myeloid precursors, megakaryocytes, and plasma cells.
Main Results:
- Normal marrows showed highest proliferation in erythroid cells (mean 90%), followed by myeloid precursors (mean 38%) and megakaryocytes (mean 14%). No Ki-67 positive plasma cells were observed.
- Erythroid cell proliferation was similar in neoplastic disorders compared to controls.
- Myeloid precursor proliferation largely overlapped normal values in myelodysplasia, chronic myeloproliferative disorders, and acute non-lymphoid leukemia (M1/M2).
- Increased Ki-67 expression in megakaryocytes, including micro-megakaryocytes, was observed in chronic myeloproliferative disorders.
- Mature myeloma showed less than 2% Ki-67 positive cells.
Conclusions:
- Erythroid and myeloid proliferative activity evaluation has limited value in differentiating myeloproliferative disorders.
- Increased megakaryocyte proliferation, detected by Ki-67, may aid in diagnosing challenging chronic myeloproliferative disorder cases.
- Ki-67 evaluation is not useful for differentiating myeloma from reactive plasmacytosis.
- Sequential double immunophenotyping is a consistent method for evaluating proliferating cells in routine bone marrow biopsies.
Abstract:
The proliferative activity of the haematopoietic and plasma cells in bone marrow was evaluated under normal and neoplastic conditions, by means of a sequential double immunostaining technique, using monoclonal antibody MIB-1 recognizing the cell proliferation-associated nuclear antigen Ki-67, and antibodies against glycophorin-C, myeloperoxidase, factor VIII-related antigen, and immunoglobulin light chains. Fifty-eight B5 fixed, paraffin-embedded bone marrow biopsies were analysed, including 11 normal controls. 10 cases of myelodysplasia, 14 cases of chronic myeloproliferative disorder, eight cases of acute non-lymphoid leukaemia, and 15 cases of myeloma. In normal marrows, the highest proliferative activity was noticed in the erythroid cells (75% to 95%; mean 90%), in comparison with myeloid precursors (15% to 80%; mean 38%), and megakaryocytes (10% to 20%; mean 14%): no Ki-67 positive plasma cells were found. In all investigated haematological disorders, the expression of MIB-1 by erythroid cells was similar to that observed in controls. Similarly, the percentage of MIB-1 + myeloid precursors in chronic myeloproliferative disorders and myelodysplasia largely overlapped the values observed in normals, and comparable values were also found in the blast cells from acute non-lymphoid leukaemia type M1 and M2. These findings suggest that the evaluation of either erythroid or myeloid proliferative activity is of little value in the differential diagnosis between these myeloproliferative disorders. By contrast, the obvious increase of Ki-67 expression of megakaryocytes in chronic myeloproliferative disorders, with labelling also of micro-megakaryocytes, might sustain the diagnosis in controversial cases. Since cases of mature myeloma showed less than 2% of Ki-67 positive cells, evaluation of proliferative activity is of no value in the differential diagnosis with reactive plasmacytosis. The sequential double immunophenotyping for Ki-67 antigen and for haematopoietic cell lineage-associated markers can be applied in a consistent manner to routine bone marrow biopsies to evaluate proliferating cells in normal and neoplastic conditions.
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