Immunophenotypic differentiation patterns of normal hematopoiesis in human bone marrow: reference patterns for age-related changes and disease-induced shifts

Cytometry B Clin Cytom. 2004 Jul;60(1):1-13. doi: 10.1002/cyto.b.20008.


Background: The abundance of monoclonal antibodies (mAb) and the routine use of quadruple stainings in flow cytometry allow stepwise analysis of bone marrow (BM) samples that are suspected for abnormal hematopoiesis. A screening phase that precedes lineage-specific classification phases should be sufficient to assess whether the BM has a normal or abnormal composition, as well as to identify the abnormal differentiation lineage.

Methods: For a quick and easy flow cytometric screening of BM samples, we selected six quadruple immunostainings that cover multiple differentiation stages of the B-cell, monocytic, granulocytic, and erythroid lineages: TdT/CD20/CD19/CD10 and CD45/CD34/CD19/CD22 for B cells, CD34/CD117/CD45/CD13.33 for precursor granulocytic and precursor monocytic cells (myelo/monoblasts), CD14/CD33/CD45/CD34 for monocytic cells, CD16/CD13/CD45/CD11b for granulocytic cells, and CD71/CD235a/CD45/CD117 for erythroid cells.

Results: The six quadruple immunostainings reveal specific staining patterns in normal BM, which allow the recognition of various subpopulations of the respective lineages. These staining patterns can be used as a frame of reference for recognition of normal and abnormal BM development. Examples of normal (age-related) variations in these otherwise stable staining patterns are presented together with several abnormal differentiation patterns.

Conclusions: Although alternative immunostainings can be used (e.g., including NK- and T-cell markers), we feel that the selected six stainings represent a comprehensive and easy screening phase for quick identification of shifts in the composition of the studied differentiation lineages, reflecting age-related changes or disease-induced BM abnormalities.

MeSH terms

  • Aging / physiology*
  • B-Lymphocytes / cytology
  • B-Lymphocytes / immunology
  • Bone Marrow / immunology
  • Bone Marrow / pathology
  • Bone Marrow / physiology*
  • Bone Marrow Cells / cytology*
  • Bone Marrow Cells / immunology
  • Cell Lineage
  • Erythroid Cells / cytology
  • Erythroid Cells / immunology
  • Flow Cytometry
  • Granulocytes / cytology
  • Granulocytes / immunology
  • Hematologic Diseases / pathology
  • Hematopoiesis*
  • Humans
  • Immunophenotyping
  • Monocytes / cytology
  • Monocytes / immunology
  • Reference Values