scholarly journals A Novel Type of Stem Cells Double-Positive for SSEA-3 and CD45 in Human Peripheral Blood

2020 ◽  
Vol 29 ◽  
pp. 096368972092357
Author(s):  
Tetsuya Sato ◽  
Shohei Wakao ◽  
Yoshihiro Kushida ◽  
Kazuki Tatsumi ◽  
Masaaki Kitada ◽  
...  
Keyword(s):  
Progenitor Cells ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Surface Marker ◽  
Cytokeratin 19 ◽  
Hematopoietic Stem ◽  
Pluripotency Markers ◽  
S1p Receptor ◽  
Muse Cells

Peripheral blood (PB) contains several types of stem/progenitor cells, including hematopoietic stem and endothelial progenitor cells. We identified a population positive for both the pluripotent surface marker SSEA-3 and leukocyte common antigen CD45 that comprises 0.04% ± 0.003% of the mononuclear cells in human PB. The average size of the SSEA-3(+)/CD45(+) cells was 10.1 ± 0.3 µm and ∼22% were positive for CD105, a mesenchymal marker; ∼85% were positive for CD19, a B cell marker; and ∼94% were positive for HLA-DR, a major histocompatibility complex class II molecule relevant to antigen presentation. These SSEA-3(+)/CD45(+) cells expressed the pluripotency markers Nanog, Oct3/4, and Sox2, as well as sphingosine-1-phosphate (S1P) receptor 2, and migrated toward S1P, although their adherence and proliferative activities in vitro were low. They expressed NeuN at 7 d, Pax7 and desmin at 7 d, and alpha-fetoprotein and cytokeratin-19 at 3 d when supplied to mouse damaged tissues of the brain, skeletal muscle and liver, respectively, suggesting the ability to spontaneously differentiate into triploblastic lineages compatible to the tissue microenvironment. Multilineage-differentiating stress enduring (Muse) cells, identified as SSEA-3(+) in tissues such as the bone marrow and organ connective tissues, express pluripotency markers, migrate to sites of damage via the S1P-S1P receptor 2 system, and differentiate spontaneously into tissue-compatible cells after homing to the damaged tissue where they participate in tissue repair. After the onset of acute myocardial infarction and stroke, patients are reported to have an increase in the number of SSEA-3(+) cells in the PB. The SSEA-3(+)/CD45(+) cells in the PB showed similarity to tissue-Muse cells, although with difference in surface marker expression and cellular properties. Thus, these findings suggest that human PB contains a subset of cells that are distinct from known stem/progenitor cells, and that CD45(+)-mononuclear cells in the PB comprise a novel subpopulation of cells that express pluripotency markers.

scholarly journals Purification and characterization of granulocytic progenitor cells (CFU- C) from human peripheral blood using immunologic surface markers

Blood ◽  
1978 ◽  
Vol 51 (1) ◽  
pp. 1-8 ◽  
Author(s):  
CM Richman ◽  
L Chess ◽  
RA Yankee
Keyword(s):  
Progenitor Cells ◽  
Peripheral Blood ◽  
B Lymphocyte ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Sheep Erythrocyte ◽  
Methyl Cellulose ◽  
Null Cell ◽  
Peripheral Blood Mononuclear

Abstract The concentration of committed granulocytic progenitor cells (CFU-C) in functionally unique subpopulations of human peripheral blood mononuclear cells has been determined by the in vitro methyl-cellulose assay. Using immunoabsorbent column chromatography and rosette-depletion techniques, we have demonstrated that CFU-C, although not present in either purified T or B lymphocyte populations, are highly concentrated in the “null” cell population, which lacks sheep erythrocyte receptors and surface immunoglobulin. Further fractionation of this null subset has demonstrated that CFU-C do not bear complement receptors, but require the presence of peripheral blood mononuclear cell feeder layers for maximum proliferation.

Low Density Granulocytes in Patients after Allogeneic Hematopoietic Stem Cells Transplantation (allo-HSCT): A Distinct Class of Neutrophils in Systemic Alloimmunity

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 4614-4614
Author(s):  
Ekaterina Mikhaltsova ◽  
Valeri G. Savchenko ◽  
Larisa A. Kuzmina ◽  
Mikhail Drokov ◽  
Vera Vasilyeva ◽  
...  
Keyword(s):  
Lupus Erythematosus ◽  
Peripheral Blood ◽  
Hematological Malignancies ◽  
Mononuclear Cells ◽  
Conflicts Of Interest ◽  
Human Peripheral Blood ◽  
Unrelated Donor ◽  
Low Density ◽  
Hematopoietic Stem

Abstract Introduction It's generally considered that all alloimmune process such as acute graft-versus host disease (aGVHD) after allo-HSCT are mostly controlled by lymphocytes. The role of neutrophils in systemic alloimmunity after allo-HSCT is still illusive. In 1987 a distinct subset of proinflammatory, low-density granulocytes (LDGs) isolated from the peripheral blood mononuclear cell fractions of patients with system lupus erythematosus has been described. There is no LDG's in healthy donors. While the origin and role of LDGs still needs to be fully characterized, we try to describe this population in patients with hematological malignancies after allo-HSCT Patients and methods. Peripheral blood samples were collected in EDTA-tubes before allo-HSCT, on day +30,+60,+90 after allo-HSCT and at day of aGVHD from 47 patients with hematological malignancies (AML=22, ALL n=17, LPD=3, MDS =2; CML=2; 17 with active disease, 30 - in CR) after allo-HSCT (from matched unrelated donor n=34, from matched related donor n=13; MAC = 13, RIC=34). Isolation of mononuclear cells from human peripheral blood was made by standard protocol using Lympholyte®-M Cell Separation Media (Cedarlane Labs). The anti-CD66b-PE (Biolegend, USA) antibodies and FSC/SSC were used to determine LDGs cells as FSChigh \SSChigh \CD66b+. 100000 of cells were analyzed on a BD FACSCanto II (Becton Dickinson, USA). Results. Results of blood evaluation of 47 patients with hematological malignancies, whose blood was examined after allo-HSCT presented in table 1. Conclusion Despite the fact that we don't get significant differences. LDG's detection in allo-HSCT patients need further investigation. Table 1. Incidence of LDG after allo-HSCT in patients with and without aGVHD Table 1. Incidence of LDG after allo-HSCT in patients with and without aGVHD Disclosures No relevant conflicts of interest to declare.

scholarly journals Nonobese Diabetic/Severe Combined Immunodeficiency (NOD/SCID) Mouse as a Model System to Study the Engraftment and Mobilization of Human Peripheral Blood Stem Cells

Blood ◽  
1998 ◽  
Vol 92 (7) ◽  
pp. 2556-2570 ◽  
Author(s):  
Johannes C.M. van der Loo ◽  
Helmut Hanenberg ◽  
Ryan J. Cooper ◽  
F.-Y. Luo ◽  
Emmanuel N. Lazaridis ◽  
...  
Keyword(s):  
Stem Cell ◽  
Progenitor Cells ◽  
Peripheral Blood ◽  
Human Peripheral Blood ◽  
Severe Combined Immunodeficiency ◽  
Human Cells ◽  
Scid Mice ◽  
Combined Immunodeficiency ◽  
Hematopoietic Stem ◽  
Nonobese Diabetic

Mobilized CD34+ cells from human peripheral blood (PB) are increasingly used for hematopoietic stem-cell transplantation. However, the mechanisms involved in the mobilization of human hematopoietic stem and progenitor cells are largely unknown. To study the mobilization of human progenitor cells in an experimental animal model in response to different treatment regimens, we injected intravenously a total of 92 immunodeficient nonobese diabetic/severe combined immunodeficiency (NOD/SCID) mice with various numbers of granulocyte colony-stimulating factor (G-CSF) –mobilized CD34+ PB cells (ranging from 2 to 50 × 106cells per animal). Engraftment of human cells was detectable for up to 6.5 months after transplantation and, depending on the number of cells injected, reached as high as 96% in the bone marrow (BM), displaying an organ-specific maturation pattern of T- and B-lymphoid and myeloid cells. Among the different mobilization regimens tested, human clonogenic cells could be mobilized from the BM into the PB (P= .019) with a high or low dose of human G-CSF, alone or in combination with human stem-cell factor (SCF), with an average increase of 4.6-fold over control. Therefore, xenotransplantation of human cells in NOD/SCID mice will provide a basis to further study the mechanisms of mobilization and the biology of the mobilized primitive human hematopoietic cell.

scholarly journals Nonobese Diabetic/Severe Combined Immunodeficiency (NOD/SCID) Mouse as a Model System to Study the Engraftment and Mobilization of Human Peripheral Blood Stem Cells

Blood ◽  
1998 ◽  
Vol 92 (7) ◽  
pp. 2556-2570 ◽  
Author(s):  
Johannes C.M. van der Loo ◽  
Helmut Hanenberg ◽  
Ryan J. Cooper ◽  
F.-Y. Luo ◽  
Emmanuel N. Lazaridis ◽  
...  
Keyword(s):  
Stem Cell ◽  
Progenitor Cells ◽  
Peripheral Blood ◽  
Human Peripheral Blood ◽  
Severe Combined Immunodeficiency ◽  
Human Cells ◽  
Scid Mice ◽  
Combined Immunodeficiency ◽  
Hematopoietic Stem ◽  
Nonobese Diabetic

Abstract Mobilized CD34+ cells from human peripheral blood (PB) are increasingly used for hematopoietic stem-cell transplantation. However, the mechanisms involved in the mobilization of human hematopoietic stem and progenitor cells are largely unknown. To study the mobilization of human progenitor cells in an experimental animal model in response to different treatment regimens, we injected intravenously a total of 92 immunodeficient nonobese diabetic/severe combined immunodeficiency (NOD/SCID) mice with various numbers of granulocyte colony-stimulating factor (G-CSF) –mobilized CD34+ PB cells (ranging from 2 to 50 × 106cells per animal). Engraftment of human cells was detectable for up to 6.5 months after transplantation and, depending on the number of cells injected, reached as high as 96% in the bone marrow (BM), displaying an organ-specific maturation pattern of T- and B-lymphoid and myeloid cells. Among the different mobilization regimens tested, human clonogenic cells could be mobilized from the BM into the PB (P= .019) with a high or low dose of human G-CSF, alone or in combination with human stem-cell factor (SCF), with an average increase of 4.6-fold over control. Therefore, xenotransplantation of human cells in NOD/SCID mice will provide a basis to further study the mechanisms of mobilization and the biology of the mobilized primitive human hematopoietic cell.

EPCs Derived from Ex Vivo Expansion of Unmobilized Human Peripheral Blood Co-Express Hematopoietic and Endothelial-Specific Markers and Can Be Derived from Culture of Highly Purified Populations of CD14-Positive Monocytes.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 2682-2682
Author(s):  
Emerson E. Sharpe ◽  
Amylynn A. Teleron ◽  
Bin Li ◽  
Pampee P. Young
Keyword(s):  
Progenitor Cells ◽  
Peripheral Blood ◽  
Ex Vivo ◽  
Human Peripheral Blood ◽  
Ex Vivo Expansion ◽  
Adherent Cells ◽  
Hematopoietic Stem ◽  
Endothelial Markers ◽  
Cell Based Therapy

Abstract An increasing amount of data has suggested a more dynamic role of vasculogenesis, whereby bone marrow (BM)-derived circulating endothelial progenitor cells (EPCs) home and contribute to new blood vessel formation during tumor growth, ischemic injury, and wound healing. EPCs can be obtained by isolating hematopoietic progenitor cells from BM or cord blood. Alternatively, ex vivo expansion of unmobilized human peripheral blood (PB) can generate adherent cells, PB-EPCs, that express endothelial markers and also, upon administration, incorporate into developing neovasculature. The relative ease of obtaining unmobilized human PB has made PB-EPCs an attractive candidate with which to develop cell based therapy to treat ischemia. In parallel with clinical trials designed to understand their therapeutic potential, there is a continued effort to better characterize the PB-EPC and understand its biology. It is currently thought that EPCs are directly derived from a CD34+/lin- faction of hematopoietic stem cells (HSCs). However, in our current study, we have confirmed prior reports that ex vivo expansion of human PB generates similar numbers of EPCs as compared to plating unfractionated human BM, which contains >50-fold higher CD34+/lin- content, suggesting that PB-EPCs may not be derived from the CD34+/lin- population. We used immunofluorescence and FACs analysis to further show that PB-EPCs not only express endothelial markers such as vWF, Vascular Endothelial Growth Factor Receptor 1 and 2 (also known as flt-1 and flk-1, respectively), VE-cadherin, UEA-1 lectin, Tie-1 and Tie-2 but also hematopoietic markers such as CD45 and CD14, a marker enriched on monocytes. To test if PB-derived CD14-positive cells can give rise to PB-EPCs, we isolated them from human PB to >98% purity and plated them on fibronectin-coated coverslips. In vitro culture of CD14-positive cells generated adherent clusters of spindle shaped cells morphologically similar to EPCs. Culture of the CD14-negative fraction failed to yield any adherent cells. After ten days, the coverslips were removed and the cells were stained with various endothelial (flk-1, vWF, uptake of DiI-AcLDL, and UEA-1 lectin) and monocyte/hematopoietic (CD14 and CD45) cell markers. In analysis of these slides, the EPCs derived from the purified CD14 fraction stained positive for all six markers. These observations suggest that PB-EPCs can differentiate from cells of the monocytic lineage in vitro without the necessity of interaction from cells contained in the CD14-negative population. Further experiments will test the possibility that monocytes may be an intermediate in the differentiation of EPCs in vivo.

scholarly journals Purification and characterization of granulocytic progenitor cells (CFU- C) from human peripheral blood using immunologic surface markers

Blood ◽  
1978 ◽  
Vol 51 (1) ◽  
pp. 1-8
Author(s):  
CM Richman ◽  
L Chess ◽  
RA Yankee
Keyword(s):  
Progenitor Cells ◽  
Peripheral Blood ◽  
B Lymphocyte ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Sheep Erythrocyte ◽  
Methyl Cellulose ◽  
Null Cell ◽  
Peripheral Blood Mononuclear

The concentration of committed granulocytic progenitor cells (CFU-C) in functionally unique subpopulations of human peripheral blood mononuclear cells has been determined by the in vitro methyl-cellulose assay. Using immunoabsorbent column chromatography and rosette-depletion techniques, we have demonstrated that CFU-C, although not present in either purified T or B lymphocyte populations, are highly concentrated in the “null” cell population, which lacks sheep erythrocyte receptors and surface immunoglobulin. Further fractionation of this null subset has demonstrated that CFU-C do not bear complement receptors, but require the presence of peripheral blood mononuclear cell feeder layers for maximum proliferation.

Cloning and Functional Analysis of the Rhesus Macaque ABCG2 Gene: Forced Expression Confers an SP Phenotype among Hematopoietic Stem Cell Progeny in Vivo.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 3219-3219 ◽  
Author(s):  
Takahiro Ueda ◽  
Sebastian Brenner ◽  
Harry Malech ◽  
Saskia Langemeijer ◽  
Martha Kirby ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Gene Transfer ◽  
Rhesus Macaque ◽  
Progenitor Cells ◽  
Peripheral Blood ◽  
Human Peripheral Blood ◽  
Hematopoietic Stem ◽  
Cell Counts ◽  
Abcg2 Gene

Abstract Hematopoietic cells can be highly enriched for repopulating ability based upon efflux of the fluorescent Hoechst 33342 dye by sorting for side population (SP) cells, a phenotype attributed to expression of ABCG2, a member of the ABC transporter superfamily. Intriguingly, murine studies suggest that forced ABCG2 expression prevents hematopoietic differentiation. We sought to determine the effects of forced expression of the ABCG2 gene in hematopoietic stem cells in the nonhuman primate model, a model with proven relevance to human hematopoiesis. We cloned the full-length rhesus ABCG2 (rh-ABCG2) cDNA using a series of primers spanning the entire sequence designed using the published human sequence. Sequence homology was greater than 96%. The rh-ABCG2 gene was then introduced into an MFGS based retroviral vector pseudotyped with the RD114 envelope. Mobilized human peripheral blood CD34-positive cells were transduced with either rh-ABCG2 or human GP91-phox vector with no other payload. All transductions were initiated with 4 x10e5 cells using X-VIVO10/1%HSA/4mM/L L-glutamine supplemented with 100ng/ml_FLT3L, 100ng/ml SCF, 100ng/ml TPO and polybrene (5 ug/ml). RD114 vector was concentrated by ultracentrifugation (83,000g 90minutes 4°C). Gene transfer rates to CFU of greater than 80% were achieved using both vectors with similar gene transfer rate estimated by flow cytometry. ABCG2-transduced human peripheral blood progenitor cells (PBPCs) acquired the SP phenotype, but showed significantly reduced growth compared to control (Day 8: cell counts 7.67+/− 2.54 vs. 17.83+/−6.64 x10e5 for ABCG2 and GP91-phox transduced cells, respectively p=0.0024, n=5). We then examined the engraftment of ABCG2-expressing stem and progenitor cells in the rhesus macaque autologous transplant model. GCSF/SCF mobilized PBPCs were collected from 2 animals and the CD34+ cells were divided and transduced with either vector and infused after lethal irradiation. In vivo marking levels post transplant measured in mononuclear cells and granulocytes from peripheral blood and bone marrow ranged initially from 0.5–4% by Realtime PCR, declined equally over time, and were similar between transduced fractions, with no discrepancy between bone marrow and peripheral blood marking. Furthermore, peripheral blood T cells, B cells and granulocytes expressed ABCG2 at levels predicted by vector copy number long term, and the differential of such cells within the SP gate matched that of the non-SP fraction demonstrating no block to differentiation in the large animal. In vitro studies showed selective protection against mitoxantrone among ABCG2-transduced rhesus PBPCs. Our results confirm the existence of rhesus-ABCG2, support its importance in conferring the SP phenotype, suggest no detrimental effect of its overexpression upon hematopoiesis, and imply a potential role for its overexpression as an in vivo selection strategy for gene therapy applications.

Hematopoietic Stem - and Progenitor Cells Are Differentially Mobilized in Response to Flt3-Ligand

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 4782-4782
Author(s):  
Evert-Jan F. M. de Kruijf ◽  
Melissa van Pel ◽  
Willem E. Fibbe
Keyword(s):  
Progenitor Cells ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Fold Increase ◽  
Hematopoietic Progenitor Cell ◽  
Tyrosine Kinase Receptor ◽  
Steady Increase ◽  
Flt3 Ligand ◽  
Hematopoietic Stem ◽  
Stem And Progenitor Cells

Abstract Flt3 is a tyrosine kinase receptor expressed mainly on primitive hematopoietic cells and its ligand, Flt3-ligand (FL), is a slowly mobilizing agent in mice when administered for 5–10 days. This provides a time-frame to study the HPC and HSC migration kinetics in detail. BALB/c mice were injected for 3, 5, 7 and 10 days with FL (10 ug/day, intraperitoneally). Mobilization of hematopoietic stem- and progenitor cells was studied using colony-forming-unit granulocyte/monocyte (CFU-GM) and cobblestone-area-forming-cell (CAFC) assays. The radioprotective capacity of mobilized peripheral blood mononuclear cells (PBMC) was studied by transplantation of 1.5 × 106 FL-mobilized PBMNC into lethally irradiated (9.5 Gy) recipients. Hematopoietic progenitor cell mobilization was detected from day 3 onwards and prolonged administration of FL showed a steady increase in mobilized progenitor cells (CFU-GM; 23.1±20.1 [PBS], 219.0±213.7 [3 days FL, n=10], 1,007.8±742 [5 days FL, n=21], 3,526.6±1,406 [7 days FL, n=10] and 19,149±2,338 [10 days FL, n=10]). Administration of FL for 10 days lead to a 5.5-fold increase in CAFC-week 4, compared to FL administration for 5 days (0.69 vs 0.13 CAFC per 105 PBMC respectively) and a 5.0-fold increase in CAFC-week 5 (0.27 vs 0.05 CAFC per 105 PBMC respectively). No CAFC week 4/5 were found in PBMC obtained from PBS-injected control mice. Transplantation of 5-day FL-mobilized PBMC did not radioprotect lethally irradiated recipients (mean survival time (MST): 66 days; MST PBS: 13 days). In contrast, transplantation of PBMC obtained from a 10-day FL mobilization regime radioprotected 100% of the recipients (Table 1). These results indicate that HPC and HSC show different mobilization kinetics in response to FL, resulting in preferential mobilization of HPC at day 5, followed by HSC mobilization at day 10. We speculate that this difference in mobilization kinetics might be due to the selective recruitment of HPC and HSC from their respective bone marrow compartments. Table 1. HPC and HSC frequencies in peripheral blood following Flt3-ligand-induced mobilization CFU-GM per ml PB CAFC-day 28 per 105cells CAFC-day 35 per 105cells % Radioprotection day 150 * fold increase compared to 5 day FL 5 days Flt3-L 1,007 (n=21) 0.126 0.054 0 (n=10) 10 days Flt3-L 19,149 (n=10) (19.0)* 0.694 (5.5)* 0.272 (5.0)* 100 (n=10)

Sign in / Sign up

Export Citation Format

Share Document