Korean J Hematol 1997; 32(1):
Published online March 31, 1997
© The Korean Society of Hematology
유세포 측정기를 이용한 정상 골수 세포의 면역표현형 분석
오은지, 김도희, 김용구, 한경자, 심상인, 정익주, 한치화, 김춘추
가톨릭대학교의과대학 임상병리학교실,
가톨릭대학교의과대학 내과학교실
Immunophenotyping of Normal Human Bone Marrow by Flow Cytometric Analysis
Background: By development of monoclonal antibodies(MoABs) directed against hematopoietic cells, flow cytometric analysis of bone marrow has become commonplace
in clinical hematology laboratories and has a major role in evaluation of lymphohematopoietic malignancies. However, little information about antigen expressions and distribution of normal human bone marrow cells has been published. Therefore, we analysed the immunophenotype of the normal human bone marrow cells to get a normal
baseline data for flow cytometric analysis.
Methods: The bone marrow aspirates of 20 healthy donors of allogeneic bone marrow transplantation were analysed using flow cytometry(Becton-Dickinson Co, USA). Seven gated region(R) were set using the forward vs. right angle light scatter(FSC/SSC) cytogram and the percentages of positive cells against 17 monoclonal antibodies were identified in these gated regions by dual parameter flow cytometry.
Results: The proportion of total CD45+ cells was 87.57±12.82%(mean±1SD, n=20) and CD45-(nucleated erythrocytes) cells was 12.43±12.82% of all nucleated bone marrow
cells. T cells were more numerous than B cells in total gates(P=.0001). T helper/inducer cells(Th) to T suppressor/cytotoxic cells(Ts) ratio was 1.38±0.53 and CD4+ cells were distributed in larger size and higher SSC fractions in FSC/SSC cytogram than CD8+ cells(P=.0001). While CD4+, 8+ cells were rarely existed, CD10+ cells were 17.63±11.43% of all nucleated bone marrow cells and they mainly distributed in granulocytic fractions. Lymphocytes represent 60% of all lymphogate cells and T cells were mostly mature cells. CD10+ cells and CD34+ cells in the R1(lymphogate) were 4.37±3.07% and 1.78±1.25%, respectively. The total CD34+ cells represent 0.88±0.5% of all nucleated bone marrow cells and 65% of them were CD34+CD33+ cells.
Conclusion: These results indicate each subpopulation of normal bone marrow reveals different regional expression from morphological estimation and these normal expressions
should be considered in flow cytometric analysis of hematologic disorders.
Keywords Human bone marrow; Flow cytometry; Immunophenotype;
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Korean J Hematol 1997; 32(1): 48-56
Published online March 31, 1997
Copyright © The Korean Society of Hematology.
유세포 측정기를 이용한 정상 골수 세포의 면역표현형 분석
오은지, 김도희, 김용구, 한경자, 심상인, 정익주, 한치화, 김춘추
가톨릭대학교의과대학 임상병리학교실,
가톨릭대학교의과대학 내과학교실
Immunophenotyping of Normal Human Bone Marrow by Flow Cytometric Analysis
Eun Jee Oh, Do Hee Kim, Yong Goo Kim, Kyungia Han, Sang In Shim, Ik Joo Jung, Chi Hwa Han, Choon Choo Kim
Department of Clinical Pathology, Internal Medicine, Catholic University Medical College, Seoul, Korea
Abstract
Background: By development of monoclonal antibodies(MoABs) directed against hematopoietic cells, flow cytometric analysis of bone marrow has become commonplace
in clinical hematology laboratories and has a major role in evaluation of lymphohematopoietic malignancies. However, little information about antigen expressions and distribution of normal human bone marrow cells has been published. Therefore, we analysed the immunophenotype of the normal human bone marrow cells to get a normal
baseline data for flow cytometric analysis.
Methods: The bone marrow aspirates of 20 healthy donors of allogeneic bone marrow transplantation were analysed using flow cytometry(Becton-Dickinson Co, USA). Seven gated region(R) were set using the forward vs. right angle light scatter(FSC/SSC) cytogram and the percentages of positive cells against 17 monoclonal antibodies were identified in these gated regions by dual parameter flow cytometry.
Results: The proportion of total CD45+ cells was 87.57±12.82%(mean±1SD, n=20) and CD45-(nucleated erythrocytes) cells was 12.43±12.82% of all nucleated bone marrow
cells. T cells were more numerous than B cells in total gates(P=.0001). T helper/inducer cells(Th) to T suppressor/cytotoxic cells(Ts) ratio was 1.38±0.53 and CD4+ cells were distributed in larger size and higher SSC fractions in FSC/SSC cytogram than CD8+ cells(P=.0001). While CD4+, 8+ cells were rarely existed, CD10+ cells were 17.63±11.43% of all nucleated bone marrow cells and they mainly distributed in granulocytic fractions. Lymphocytes represent 60% of all lymphogate cells and T cells were mostly mature cells. CD10+ cells and CD34+ cells in the R1(lymphogate) were 4.37±3.07% and 1.78±1.25%, respectively. The total CD34+ cells represent 0.88±0.5% of all nucleated bone marrow cells and 65% of them were CD34+CD33+ cells.
Conclusion: These results indicate each subpopulation of normal bone marrow reveals different regional expression from morphological estimation and these normal expressions
should be considered in flow cytometric analysis of hematologic disorders.
Keywords: Human bone marrow, Flow cytometry, Immunophenotype,
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