Delineation among eight major hematopoietic subsets in murine bone marrow using a two-color flow cytometric technique.
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.
Department of Pathology, University of Iowa College of Medicine, Iowa City, Iowa 52242, USA. annette-schlueter@uiowa.edu
A J
AJ
Schlueter
S K
SK
Bhatia
X
X
Li
L T
LT
Tygrett
Y
Y
Yamashita
P
P
de Vries
T J
TJ
Waldschmidt
D000818:Animals; D000950:Antigens, Ly / Q000032:analysis; D001854:Bone Marrow Cells / Q000737:chemistry / Q000145:classification / Q000166:cytology*; D003713:Dendritic Cells / Q000737:chemistry / Q000145:classification / Q000166:cytology; D005434:Flow Cytometry / Q000379:methods*; D006410:Hematopoiesis / Q000502:physiology; D006412:Hematopoietic Stem Cells / Q000737:chemistry / Q000145:classification / Q000166:cytology*; D017493:Leukocyte Common Antigens / Q000032:analysis; D008565:Membrane Proteins / Q000032:analysis; D051379:Mice; D008807:Mice, Inbred BALB C; D013047:Specific Pathogen-Free Organisms
D000950:Antigens, Ly; C074650:Ly6a protein, mouse; D008565:Membrane Proteins; D017493:Leukocyte Common Antigens