scholarly journals Characterization and functional analysis of adult human bone marrow cell subsets in relation to B-lymphoid development

Blood ◽  
1993 ◽  
Vol 82 (2) ◽  
pp. 417-429 ◽  
Author(s):  
S Pontvert-Delucq ◽  
J Breton-Gorius ◽  
C Schmitt ◽  
C Baillou ◽  
J Guichard ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Mononuclear Cells ◽  
Marrow Cell ◽  
Light And Electron Microscopy ◽  
Small Cells ◽  
Multiparameter Flow Cytometry ◽  
Hla Dr ◽  
B Lineage ◽  
B Lymphoid ◽  
High Level

Abstract To study the frontiers between pluripotent stem cells and committed progenitors and to further define the B-cell pathway in adult bone marrow (BM), CD34+ subpopulations and CD34- B-lineage cells were analyzed by multiparameter flow cytometry, studied by light and electron microscopy, and in short-term and long-term cultures (LTC). While the total CD34+ cells represent 4.9% +/- 0.8 of BM mononuclear cells within the lymphoid-blast window, 73.8 +/- 3.5%, 14.4 +/- 1.8% and 8.8 +/- 2.9% of them were CD34+ CD10- CD19-, CD34+ CD10+ CD19+, and CD34+ CD10+ CD19-, respectively. CD34+ CD10+ CD19+ cells represent a smal homogeneous TdT4 c micro-blast population. Although expressing CD38 and high level of HLA-DR antigens, like myeloid committed progenitors, they did not generate LTC, myeloid, and T lymphoid colonies suggesting that the CD34+ CD10+ CD19+ population represents exclusively B lymphoid committed progenitors. By contrast, all myeloid progenitors and LTC-initiating cells were found in the CD34+ CD10- CD19- cell fraction. This fraction appeared more heterogeneous and contained CD38- HLA-DRlow small cells, larger blasts, and promonocyte-like cells exhibiting small peroxidase-positive granules. Interestingly, CD10 was also present on CD34+ CD19- cells. This population mainly coexpressed CD33 and gave rise to macrophagic colonies.

scholarly journals Characterization and functional analysis of adult human bone marrow cell subsets in relation to B-lymphoid development

Blood ◽  
1993 ◽  
Vol 82 (2) ◽  
pp. 417-429 ◽  
Author(s):  
S Pontvert-Delucq ◽  
J Breton-Gorius ◽  
C Schmitt ◽  
C Baillou ◽  
J Guichard ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Mononuclear Cells ◽  
Marrow Cell ◽  
Light And Electron Microscopy ◽  
Small Cells ◽  
Multiparameter Flow Cytometry ◽  
Hla Dr ◽  
B Lineage ◽  
B Lymphoid ◽  
High Level

To study the frontiers between pluripotent stem cells and committed progenitors and to further define the B-cell pathway in adult bone marrow (BM), CD34+ subpopulations and CD34- B-lineage cells were analyzed by multiparameter flow cytometry, studied by light and electron microscopy, and in short-term and long-term cultures (LTC). While the total CD34+ cells represent 4.9% +/- 0.8 of BM mononuclear cells within the lymphoid-blast window, 73.8 +/- 3.5%, 14.4 +/- 1.8% and 8.8 +/- 2.9% of them were CD34+ CD10- CD19-, CD34+ CD10+ CD19+, and CD34+ CD10+ CD19-, respectively. CD34+ CD10+ CD19+ cells represent a smal homogeneous TdT4 c micro-blast population. Although expressing CD38 and high level of HLA-DR antigens, like myeloid committed progenitors, they did not generate LTC, myeloid, and T lymphoid colonies suggesting that the CD34+ CD10+ CD19+ population represents exclusively B lymphoid committed progenitors. By contrast, all myeloid progenitors and LTC-initiating cells were found in the CD34+ CD10- CD19- cell fraction. This fraction appeared more heterogeneous and contained CD38- HLA-DRlow small cells, larger blasts, and promonocyte-like cells exhibiting small peroxidase-positive granules. Interestingly, CD10 was also present on CD34+ CD19- cells. This population mainly coexpressed CD33 and gave rise to macrophagic colonies.

scholarly journals Normal human bone marrow precursors that express terminal deoxynucleotidyl transferase include T-cell precursors and possible lymphoid stem cells

Blood ◽  
1991 ◽  
Vol 77 (8) ◽  
pp. 1681-1690
Author(s):  
SD Gore ◽  
MB Kastan ◽  
CI Civin
Keyword(s):  
Bone Marrow ◽  
T Cell ◽  
Mononuclear Cells ◽  
Cell Receptor ◽  
Small Population ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Immune Adherence ◽  
Hla Dr ◽  
Major Subset ◽  
B Lymphoid

To compare the differentiation of early B- and T-lymphoid precursors, we have used immune adherence combined with analytical flow cytometric techniques to enrich and characterize subsets of the small population of bone marrow mononuclear cells that express the enzyme terminal deoxynucleotidyl transferase (TdT) but lack the CD19 B-lymphoid marker. Two percent to five percent of bone marrow TdT + mononuclear cells belong to the T-lymphoid lineage by virtue of expression of CD7 or CD5. Three-color immunofluorescence studies showed that, like early B- lymphoid precursors, most bone marrow TdT + T cells express HLA-DR and the progenitor cell antigen CD34, and about half express CD10. All CD5 + TdT + cells express surface CD3 and T-cell receptor alpha, beta, while a subset of CD7 + TdT + cells lack these “mature” T cell features. CD2 is low or absent on CD5 + TdT + cells. Examination of isolated CD34 + cells showed that approximately 70% of CD34 + TdT + cells expressed neither CD19, CD22, CD7, nor CD5, and 15% to 50% also lacked CD10. Thus, a major subset of CD34 + TdT + cells lack lineage- specific surface antigens. TdT expression may be the earliest available marker of lymphoid differentiation, and CD34 + TdT + cells are likely to include progenitor cells for both the B and T lineages.

scholarly journals Normal human bone marrow precursors that express terminal deoxynucleotidyl transferase include T-cell precursors and possible lymphoid stem cells

Blood ◽  
1991 ◽  
Vol 77 (8) ◽  
pp. 1681-1690 ◽  
Author(s):  
SD Gore ◽  
MB Kastan ◽  
CI Civin
Keyword(s):  
Bone Marrow ◽  
T Cell ◽  
Mononuclear Cells ◽  
Cell Receptor ◽  
Small Population ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Immune Adherence ◽  
Hla Dr ◽  
Major Subset ◽  
B Lymphoid

Abstract To compare the differentiation of early B- and T-lymphoid precursors, we have used immune adherence combined with analytical flow cytometric techniques to enrich and characterize subsets of the small population of bone marrow mononuclear cells that express the enzyme terminal deoxynucleotidyl transferase (TdT) but lack the CD19 B-lymphoid marker. Two percent to five percent of bone marrow TdT + mononuclear cells belong to the T-lymphoid lineage by virtue of expression of CD7 or CD5. Three-color immunofluorescence studies showed that, like early B- lymphoid precursors, most bone marrow TdT + T cells express HLA-DR and the progenitor cell antigen CD34, and about half express CD10. All CD5 + TdT + cells express surface CD3 and T-cell receptor alpha, beta, while a subset of CD7 + TdT + cells lack these “mature” T cell features. CD2 is low or absent on CD5 + TdT + cells. Examination of isolated CD34 + cells showed that approximately 70% of CD34 + TdT + cells expressed neither CD19, CD22, CD7, nor CD5, and 15% to 50% also lacked CD10. Thus, a major subset of CD34 + TdT + cells lack lineage- specific surface antigens. TdT expression may be the earliest available marker of lymphoid differentiation, and CD34 + TdT + cells are likely to include progenitor cells for both the B and T lineages.

Immunophenotypic Study of Basophils by Multiparameter Flow Cytometry

2008 ◽  
Vol 132 (5) ◽  
pp. 813-819
Author(s):  
Xiaohong Han ◽  
Jeffrey L. Jorgensen ◽  
Archana Brahmandam ◽  
Ellen Schlette ◽  
Yang O. Huh ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
Chronic Myelogenous Leukemia ◽  
Peripheral Blood ◽  
Myelogenous Leukemia ◽  
Multiparameter Flow Cytometry ◽  
Color Flow ◽  
Aberrant Expression ◽  
Flow Cytometric ◽  
Hla Dr

Abstract Context.—The immunophenotypic profile of basophils is not yet fully established, and the immunophenotypic changes in chronic myelogenous leukemia are not fully characterized. Objective.—To establish a comprehensive immunophenotypic spectrum of normal basophils and to assess the range of immunophenotypic aberrations of basophils in chronic myelogenous leukemia. Design.—Using 4-color flow cytometry, we compared the immunophenotypic profile of basophils in peripheral blood or bone marrow samples from 20 patients with no evidence of neoplasia to basophils from 15 patients with chronic myelogenous leukemia. Results.—Basophils in control cases were all positive for CD9, CD13, CD22, CD25 (dim), CD33, CD36, CD38 (bright), CD45 (dimmer than lymphocytes and brighter than myeloblasts), and CD123 (bright), and were negative for CD19, CD34, CD64, CD117, and HLA-DR. Basophils in all chronic myelogenous leukemia patients possessed 1 to 5 immunophenotypic aberrancies. The most common aberrancies were underexpression of CD38, followed by aberrant expression of CD64 and underexpression of CD123. CD34 and CD117 were present in cases with basophilic precursors. Myeloblasts showed a distinct immunophenotypic profile, as they typically expressed CD34 and CD117, showed dimmer expression (compared with basophils) of CD38, CD45, and CD123, and lacked expression of CD22. Conclusions.—Flow cytometric immunophenotyping can identify immunophenotypic aberrations of basophils in chronic myelogenous leukemia, and discriminate basophils from myeloblasts.

scholarly journals Biology of human megakaryocyte factor V

Blood ◽  
1986 ◽  
Vol 67 (6) ◽  
pp. 1639-1648
Author(s):  
AM Gewirtz ◽  
M Keefer ◽  
K Doshi ◽  
AE Annamalai ◽  
HC Chiu ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Mononuclear Cells ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Small Cells ◽  
Factor V ◽  
Plasma Clot ◽  
Detection Techniques ◽  
Immunochemical Detection ◽  
Small Bone

To learn more about human megakaryocyte coagulation cofactor V (FV), we studied the expression of this protein in normal bone marrow megakaryocytes and in megakaryocytes cloned from their colony-forming unit in FV-depleted plasma clot cultures. Mouse monoclonal antibodies directed against either the light chain or an activation peptide of human FV and a rabbit polyclonal, monospecific FV antiserum were used as probes for these experiments in conjunction with a variety of immunochemical detection techniques. All morphologically recognizable megakaryocytes were shown to contain FV. The origin of this protein appeared to be both from FV bound to the cell as well as from endogenous FV in the majority of cells examined. The existence of a population of small bone marrow mononuclear cells that simultaneously expressed platelet glycoproteins and FV was also noted. Such cells represented approximately 70% of all small cells positive for platelet glycoproteins. In contrast, only about 40% of megakaryocyte colonies cloned in FV-deficient medium contained cells with immunochemically detectable FV. FV expression was most clearly demonstrated in large cells in the colonies, whereas smaller, presumably less mature cells labeled weakly or not at all. Synthesis of FV by human megakaryocytes was documented using elutriation-enriched cells incubated in 35S- methionine-containing medium. Megakaryocyte lysates and medium conditioned by these cells were subjected to immunoaffinity column purification. Column eluates analyzed by sodium dodecyl sulfate- polyacrylamide gel electrophoresis and autoradiography revealed radioactive bands comigrating with the heavy and light chains of thrombin-activated FV. These studies suggest that human megakaryocytes both bind and synthesize FV. Expression of these traits appears to be related to cell maturation, with binding ability appearing earlier than the ability to synthesize this protein. Finally, although the ability to bind FV appears to be universal among megakaryocytes, our culture data suggest that synthesis may be a restricted, or constitutively expressed property of these cells.

scholarly journals Comparative Dose of Intracarotid Autologous Bone Marrow Mononuclear Therapy in Chronic Ischemic Stroke in Rats

2021 ◽  
Vol 9 (A) ◽  
pp. 233-243
Author(s):  
Feda Makkiyah ◽  
Wismaji Sadewo ◽  
Rahmah Nurrizka
Keyword(s):  
Bone Marrow ◽  
Body Weight ◽  
Ischemic Stroke ◽  
Test Scores ◽  
Mononuclear Cells ◽  
Marrow Cell ◽  
Optimum Dose ◽  
Infarct Zone ◽  
Positive Effects ◽  
Cylinder Test

Research on chronic ischemic stroke is limited. One of the more promising approaches showing positive effects in the acute stage is mononuclear bone marrow cell therapy. This research may be the first which presents data about the optimum dose of bone marrow mononuclear cells (BM-MNCs) for chronic ischemic stroke in rats and discusses factors influencing recovery in the chronic stage. We performed temporary middle cerebral artery occlusion (MCAO)  procedures on the rats which were then randomly assigned to one of two experimental groups in which they were given either low or high doses of autologous BM-MNCs  (5 million or 10 million cells per kg body weight). Rat brains were fixed for HE, CD31, and doublecortin staining for analysis of the effects. Rat behavior was assessed weekly using the cylinder test and a modified neurological severity score (NSS) test. In the four weeks prior to administration of BM-MNC, cylinder test scores improved to near normal, and NSS test scores improved moderately. The infarct zone decreased significantly (p <0,01),  there was an improvement in angiogenesis (p = 0.1590) and a significant improvement in neurogenesis (p <0,01). Reduction of the infarct zone was associated with a higher dose whereas both higher and lower doses were found to have a similar effect on improving angiogenesis, and neurogenesis. Recovery was superior after twelve weeks compared with the recovery assessment at eight weeks. In conclusion, a dose of 10 million cells was more effective than a dose of 5 million cells per kg body weight for reducing the infarct zone and ameliorating neurogenesis. There was an improvement of histopathological parameters associated with the longer infarct period.

Abstract 697: Mobilization of Oct-4 + Bone Marrow-Derived Very Small Embryonic-Like Stem Cells in Patients with Acute Myocardial Infarction

Circulation ◽  
2007 ◽  
Vol 116 (suppl_16) ◽  
Author(s):  
Wojciech Wojakowski ◽  
Magda Kucia ◽  
Boguslaw Machalinski ◽  
Edyta Paczkowska ◽  
Joanna Ciosek ◽  
...  
Keyword(s):  
Myocardial Infarction ◽  
Acute Myocardial Infarction ◽  
Stem Cells ◽  
Bone Marrow ◽  
Pluripotent Stem Cells ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Small Cells ◽  
Acute Mi

Bone marrow-derived CD34 + CXCR4 + progenitor cells are mobilized into peripheral blood early in acute myocardial infarction (MI). Adult murine bone marrow contains population of small CD34 + lin − CD45 − CXCR4 + cells expressing markers of pluripotent stem cells (PSC) SSEA, Oct-4 and Nanog. This population of very small embryonic-like cells (VSEL) has unique morphology (small size 2– 4 μm, large nucleus, euchromatin) and capability to form embrioid bodies (EB). Murine EB-derived cells can in vitro differentiate into cells from all three germ layers including cardiomyocytes. We hypothesized that in patients with acute MI small cells expressing the VSEL immunophenotype and PSC markers are present in bone marrow and mobilized into peripheral blood. Blood samples (20 mL) from 18 patients with acute MI were obtained after 12 hours, 2 and 5 days after symptoms onset. Bone marrow samples (20 mL) were obtained from 2 patients with acute MI and 3 healthy volunteers. Mononuclear cells were isolated using hypotonic lysis and samples were analyzed by FACS. Mobilization of following cell populations was confirmed: hematopoietic lin − CD45 + CXCR4 + , lin − CD45 + CD133 + , lin − CD45 + CD34 + and non-hematopoietic (VSEL) lin − CD45 − CXCR4 + , lin − CD45 − CD133 + , lin − CD45 − CD34 + . Analysis of the cell number using lymphocyte gate showed more significant increase of CD45 + (hematopoietic) populations of lin − CD34 + , lin − CD133 + and lin − CXCR4 + cells. After gating for small events (VSEL size range) we found more significant mobilization of small, non-hematopoietic populations of lin − CD34 + , lin − CD133 + and lin − CXCR4 + cells (Table ). The expression of PSC markers (Oct-4, Nanog, SSEA-1) in VSEL was confirmed using real-time RT-PCR. Conclusion: We report for the first time that acute MI is associated with mobilization of non-hematopoietic VSELs expressing pluripotent stem cells markers.

scholarly journals Identification of novel B-lineage cells in human fetal bone marrow that coexpress CD7 [see comments]

Blood ◽  
1991 ◽  
Vol 77 (1) ◽  
pp. 64-68 ◽  
Author(s):  
ER Grumayer ◽  
F Griesinger ◽  
DS Hummell ◽  
RD Brunning ◽  
JH Kersey
Keyword(s):  
Bone Marrow ◽  
Cell Sorting ◽  
B Cell Development ◽  
Multiparameter Flow Cytometry ◽  
Fetal Bone ◽  
Lymphoid Development ◽  
Alternate Pathway ◽  
Lineage Markers ◽  
B Lineage

Abstract In the present study we used multiparameter flow cytometry and cell sorting to evaluate fetal bone marrow, a rich source of cells early in lymphoid development. We found CD7 to be expressed on a subset of CD19+ cells, including some that had matured to cytoplasmic mu+ (pre-B) and surface mu+ (B) cells. In addition, a less mature CD7+19+ population was characterized as mu- and CD34+/-. The CD7+19+ population was clearly distinct from the mature T cells. The CD7+19+ cells were negative for nuclear TdT in contrast to CD7–19+ cells, which frequently contained TdT. CD10, which is coexpressed on the cell surface of more than 90% of CD19+ lymphocytes, was detected in a minority of CD7+19+ lymphocytes. The CD7+19+34+ cell population may be B-lineage committed, or may represent uncommitted lymphoid precursors. The biologic role of the expression of CD7 on immature and mature cells, including those of the B lineage, may indicate (1) the presence of CD7+19+ lymphoid precursor cells and/or (2) an alternate pathway of B-cell development, in which cells coexpress CD7 with other B-lineage markers.

scholarly journals Flow cytometric analysis of human bone marrow. II. Normal B lymphocyte development

Blood ◽  
1987 ◽  
Vol 70 (5) ◽  
pp. 1316-1324 ◽  
Author(s):  
MR Loken ◽  
VO Shah ◽  
KL Dattilio ◽  
CI Civin
Keyword(s):  
Bone Marrow ◽  
Cell Surface ◽  
B Lymphocyte ◽  
Human Bone ◽  
Lymphocyte Development ◽  
Hla Dq ◽  
B Lymphocyte Development ◽  
B Lineage ◽  
B Lymphoid ◽  
B Lineage Cells

Abstract A panel of B lymphoid-reactive monoclonal antibodies was used to analyze the phenotypic changes that accompany B lymphocyte development in normal human bone marrow. The B lymphoid cells were identified using light scattering and the expression of CD19 on a flow cytometer. Quantitative three-color immunofluorescence was then used to correlate other cell surface antigens on these cells identified as B lymphoid in normal marrow. CD10 and CD20 identified almost exclusive populations and provided a convenient means of discriminating between the less and more mature B lineage cells. The CD10+ cells could be further subdivided using CD34. The population of CD19+, CD10+, CD34+ cells comprised only 0.6% of marrow cells, but these contained the majority of terminal deoxynucleotidyl transferase (TdT+) cells. In the assessment of class II antigens, HLA-DR was expressed on all B lineage cells whereas HLA-DP preceded HLA-DQ in appearance during the developmental process. Among the later antigens expressed on B lineage cells, cell surface IgM, CD20, and HLA-DQ were expressed at essentially the same time. Cell surface CD10 was lost at the time when CD21 and CD22 were acquired on the cell surface. These data illustrate that multiparameter flow cytometry can be used to define a continuous progression of stages of B lymphocyte development based on cell surface antigen expression even though these cells represent a minor fraction of normal marrow cells.

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