Open AccessDelineation among eight major hematopoietic subsets in murine bone marrow using a two‐color flow cytometric technique
Author(s) -
Schlueter Annette J.,
Bhatia Sudershan K.,
Li Xiang,
Tygrett Lorraine T.,
Yamashita Yoshio,
de Vries Peter,
Waldschmidt Thomas J.
Publication year - 2001
Publication title -
cytometry
Language(s) - English
Resource type - Journals
eISSN - 1097-0320
pISSN - 0196-4763
DOI - 10.1002/1097-0320(20010401)43:4<297::aid-cyto1062>3.0.co;2-9
Subject(s) - bone marrow , flow cytometry , biology , haematopoiesis , population , phenotype , immunology , immunophenotyping , cytometry , microbiology and biotechnology , progenitor cell , stem cell , pathology , genetics , medicine , gene , environmental health
Background Many methods have been developed specifically for identifying hematopoietic progenitor cells in murine bone marrow, but few methods allow rapid identification of multiple bone marrow populations. We describe a new, simple method for identifying simultaneously eight populations in murine bone marrow with two‐color flow cytometry and phenotypically define these populations. Methods Bone marrow was stained with anti‐Ly‐6C and anti‐B220 (CD45R) in one fluorochrome and wheat germ agglutinin (WGA) in another fluorochrome. The eight populations identified in this way were defined further primarily by four‐color flow cytometry. Results Six of the eight populations were characterized phenotypically as containing erythroid, granulocytic, mast, early B, mature B, and stem cell populations. Two additional populations with phenotypic characteristics of partially differentiated precursor cells also were identified. One population was Ly‐6C/B220+ and WGA−. It also expressed markers associated with early B, T, and/or dendritic cell differentiation. The second population was Ly‐6C hi WGA hi Mac‐1+ and was negative for numerous other lineage‐specific and precursor markers. Its morphology suggested monocytic differentiative potential. Conclusions A two‐color flow cytometric assay profiles six bone marrow populations with identifiable phenotypes and two additional unique, putative hematopoietic precursor populations. Cytometry 43:297–307, 2001. © 2001 Wiley‐Liss, Inc.