Different Adhesive Characteristics and VLA-4 Expression of CD34+ Progenitors in G0/G1 Versus S+G2/M Phases of the Cell Cycle

Blood ◽  
1998 ◽  
Vol 92 (3) ◽  
pp. 842-848 ◽  
Author(s):  
Miki Yamaguchi ◽  
Kenji Ikebuchi ◽  
Fumiya Hirayama ◽  
Norihiro Sato ◽  
Yuko Mogi ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Stromal Cell ◽  
Cell Monolayer ◽  
G1 Phase ◽  
Healthy Donors ◽  
Cd34 Cells ◽  
M Phase ◽  
American Society ◽  
Cell Cycle Status ◽  
Stimulating Factor

We identified the cell cycle status of CD34+ cells of steady-state bone marrow (BM) and peripheral blood (PB) obtained from healthy volunteers, and those of apherasis PB samples collected from healthy donors who had been administered granulocyte colony-stimulating factor (G-CSF). More than 10% of CD34+ cells in BM were in S+G2/M phase. In contrast, regardless of whether G-CSF treatment was performed, less than 2% of CD34+ cells in PB were cycling. BM CD34+ cells showed greater VLA-4 expression and adherence to stromal cells than PB CD34+cells. In addition, when cycling and dormant BM CD34+cells were analyzed separately, the cells in S+G2/M phase expressed more VLA-4 and adhered to the stromal cell monolayer more efficiently than the cells in G0/G1 phase. Furthermore, this adhesion of CD34+ cells to the stromal cell layer was almost completely inhibited by anti-VLA-4 antibody. Taken together, these results suggest that CD34+ progenitors in G0/G1 phase of the cell cycle differ from those in S+G2/M phase in adhesiveness mediated by VLA-4 in the hematopoietic microenvironment. © 1998 by The American Society of Hematology.

Different Adhesive Characteristics and VLA-4 Expression of CD34+ Progenitors in G0/G1 Versus S+G2/M Phases of the Cell Cycle

Blood ◽  
1998 ◽  
Vol 92 (3) ◽  
pp. 842-848 ◽  
Author(s):  
Miki Yamaguchi ◽  
Kenji Ikebuchi ◽  
Fumiya Hirayama ◽  
Norihiro Sato ◽  
Yuko Mogi ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Stromal Cell ◽  
Cell Monolayer ◽  
G1 Phase ◽  
Healthy Donors ◽  
Cd34 Cells ◽  
M Phase ◽  
American Society ◽  
Cell Cycle Status ◽  
Stimulating Factor

Abstract We identified the cell cycle status of CD34+ cells of steady-state bone marrow (BM) and peripheral blood (PB) obtained from healthy volunteers, and those of apherasis PB samples collected from healthy donors who had been administered granulocyte colony-stimulating factor (G-CSF). More than 10% of CD34+ cells in BM were in S+G2/M phase. In contrast, regardless of whether G-CSF treatment was performed, less than 2% of CD34+ cells in PB were cycling. BM CD34+ cells showed greater VLA-4 expression and adherence to stromal cells than PB CD34+cells. In addition, when cycling and dormant BM CD34+cells were analyzed separately, the cells in S+G2/M phase expressed more VLA-4 and adhered to the stromal cell monolayer more efficiently than the cells in G0/G1 phase. Furthermore, this adhesion of CD34+ cells to the stromal cell layer was almost completely inhibited by anti-VLA-4 antibody. Taken together, these results suggest that CD34+ progenitors in G0/G1 phase of the cell cycle differ from those in S+G2/M phase in adhesiveness mediated by VLA-4 in the hematopoietic microenvironment. © 1998 by The American Society of Hematology.

scholarly journals Peripheral blood CD34+ cells differ from bone marrow CD34+ cells in Thy- 1 expression and cell cycle status in nonhuman primates mobilized or not mobilized with granulocyte colony-stimulating factor and/or stem cell factor

Blood ◽  
1996 ◽  
Vol 87 (4) ◽  
pp. 1644-1653 ◽  
Author(s):  
RE Donahue ◽  
MR Kirby ◽  
ME Metzger ◽  
BA Agricola ◽  
SE Sellers ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Stem Cell Factor ◽  
Nonhuman Primates ◽  
Mononuclear Cells ◽  
Cytokine Therapy ◽  
Granulocyte Colony Stimulating Factor ◽  
Cd34 Cells ◽  
M Phase ◽  
Cell Cycle Status ◽  
Stimulating Factor

Granulocyte colony-stimulating factor (G-CSF) and stem cell factor (SCF) have been shown to stimulate the circulation of hematopoietic progenitor cells in both mice and nonhuman primates. We evaluated the immunophenotype and cell cycle status of CD34+ cells isolated from the bone marrow (BM) and leukapheresis product of cytokine-mobilized nonhuman primates. CD34+ cells were isolated from rhesus macaques that had received no cytokine therapy, 100 micrograms/kg/d G-CSF, 200 micrograms/kg/d SCF, or a combination of both 100 micrograms/kg/d G-CSF and 200 micrograms/kg/d SCF as a subcutaneous injection for 5 days. BM was aspirated before (day 0) and on the last day (day 5) of cytokine administration. On days 4 and 5, peripheral blood (PB) mononuclear cells were collected using a novel method of leukapheresis. Threefold more PB mononuclear cells were collected from animals receiving G-CSF alone or G-CSF and SCF than from animals that had received either SCF alone or no cytokine therapy. CD34+ cells were positively selected using an immunoadsorptive system from the BM, PB, and/or leukapheresis product. Threefold and 10-fold more CD34+ cells were isolated from the leukapheresis product of animals receiving G-CSF or G-CSF and SCF, respectively, than from animals receiving no cytokine therapy or SCF alone. The isolated CD34+ cells were immunophenotyped using CD34- allophycocyanin, CD38-fluorescein isothiocyanate, and Thy-1- phycoerythrin. These cells were later stained with 4′, 6-diamidino-2- phenylindole for simultaneous DNA analysis and immunophenotyping. BM- derived CD34+ cells did not differ significantly in cell cycle status and Thy-1 or CD38 phenotype before or after G-CSF and/or SCF administration. Similarly, CD34+ cells isolated from the leukapheresis product did not differ significantly in immunophenotype or cell cycle status before or after G-CSF and/or SCF administration. However, there were consistent differences in both immunophenotype and cell cycle status between BM- and PB-derived CD34+ cells. CD34+ cells isolated from the PB consistently had a smaller percentage of cells in the S+G2/M phase of the cell cycle and had a higher percentage of cells expressing Thy-1 than did CD34+ cells isolated from the BM. A greater proportion of PB-derived CD34+ cells were in the S+G2/M phase of the cell cycle after culture in media supplemented with interleukin-6 and SCF, However, culturing decreased the proportion of CD34+ cells expressing Thy-1.

The effect of pre-operative autologous blood donation on bone marrow hematopoietic functions in rabbits after hepatectomy

2021 ◽  
Vol 22 ◽  
Author(s):  
Xiao-Fang Zhou ◽  
Yang Liu ◽  
Jia-Ming Xu ◽  
Jin-Huo Wang ◽  
Zhen-Zhou Li ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Bone Marrow ◽  
Telomere Length ◽  
Blood Donation ◽  
Autologous Blood ◽  
G1 Phase ◽  
Control Group ◽  
Autologous Blood Donation ◽  
Cd34 Cells ◽  
Cell Cycle Status

Background: Pre-operative autologous blood donation (PABD) is one of the most widely distributed autologous blood donation means, which has positive effect on erythropoiesis. However, whether PABD can stimulate the bone marrow hematopoiesis after hepatectomy have not been reported. Methods: Totally 80 New Zealand rabbits were randomly divided into 4 groups that included control group, surgery group, hemodilutional autotransfusion (HA) group and PABD group. Automatic reticulocyte examination was performed to detect the content of reticulocyte and immature reticulocyte fractions (IRF). Flow cytometric analysis was employed to monitor the level of CD34+ cells and the cell cycle status. Southern blotting was conducted to determine the telomere length of CD34+ cells. Results: The content of high fluorescence reticulocytes (HFR) and IRF was decreased at 6 h and 24 h after autotransfusion. However, the level of CD34+ cells was upregulated after PABD. Cell cycle status analysis revealed that majority of the CD34+ cells in HA and PABD group were maintained in G0/G1 phase. The telomere length in HA and PABD group was shorten than that of control group and surgery group. Conclusion: PABD could promote the bone marrow hematopoietic functions in rabbits after hepatectomy via stimulating proliferation of CD34+ cells and shortening the telomere length of CD34+ cells, but the content of HFR was not increased immediately because of the stuck of CD34+ cells in G0/G1 phase.

scholarly journals Myeloid differentiation of purified CD34+ cells after stimulation with recombinant human granulocyte-monocyte colony-stimulating factor (CSF), granulocyte-CSF, monocyte-CSF, and interleukin-3

Blood ◽  
1991 ◽  
Vol 78 (12) ◽  
pp. 3192-3199 ◽  
Author(s):  
T Egeland ◽  
R Steen ◽  
H Quarsten ◽  
G Gaudernack ◽  
YC Yang ◽  
...  
Keyword(s):  
Myeloid Differentiation ◽  
Cell Multiplication ◽  
Colony Stimulating Factor ◽  
Interleukin 3 ◽  
Gm Csf ◽  
Healthy Donors ◽  
Cd34 Cells ◽  
Proliferation And Differentiation ◽  
Stimulating Factor

Abstract CD34+ cells isolated from bone marrow or umbilical cord blood from healthy donors were studied for proliferation and differentiation in liquid cultures in the presence of recombinant human granulocyte- monocyte colony-stimulating factor (GM-CSF), granulocyte CSF (G-CSF), monocyte CSF (M-CSF), and interleukin-3 (IL-3), followed by immunophenotyping for myeloid and myeloid-associated cell surface markers. IL-3, either alone or together with GM-CSF, G-CSF, or M-CSF, induced, on average, 50-fold cell multiplication, GM-CSF five fold to 10-fold, and G-CSF and M-CSF less than fivefold. Cells from cultures stimulated with GM-CSF, G-CSF, or M-CSF alone contained cells with a “broad” myeloid profile, “broader” than observed in cultures with IL-3. However, since IL-3 induced rapid cell multiplication, high numbers of cells expressing early (CD13, CD33) and late myeloid markers (CD14, CD15) were recovered. The presence of other CSFs together with IL-3 did not alter the IL-3-induced effect on the cells. When 5,000 CD34+ cells were cultured with IL-3 alone, the cultures still contained 2,000 to 5,000 CD34+ cells after 14 days of culture, while cells cultured with GM-CSF, G-CSF, or M-CSF contained less than 1,000 CD34+ cells. Furthermore, 1,000 to 3,000 cells were positive for the megakaryocytic lineage marker CD41b after cultures with GM-CSF or IL-3, while cultures with G-CSF or M-CSF did not contain detectable numbers of CD41b+ cells. Finally, erythroid cells could also be generated from purified CD34+ cells. The results show that IL-3 and GM-CSF can induce rapid proliferation of purified CD34+ cells in vitro with differentiation to multiple myeloid lineages, while certain subsets maintain expression of CD34.

scholarly journals Fluctuation in radioresponse of HeLa cells during the cell cycle evaluated based on micronucleus frequency

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Hiroaki Shimono ◽  
Atsushi Kaida ◽  
Hisao Homma ◽  
Hitomi Nojima ◽  
Yusuke Onozato ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Hela Cells ◽  
Time Lapse ◽  
S Phase ◽  
G1 Phase ◽  
M Phase ◽  
G2 Phase ◽  
The Difference ◽  
Time Lapse Imaging ◽  
First Time

AbstractIn this study, we examined the fluctuation in radioresponse of HeLa cells during the cell cycle. For this purpose, we used HeLa cells expressing two types of fluorescent ubiquitination-based cell cycle indicators (Fucci), HeLa-Fucci (CA)2 and HeLa-Fucci (SA), and combined this approach with the micronucleus (MN) assay to assess radioresponse. The Fucci system distinguishes cell cycle phases based on the colour of fluorescence and cell morphology under live conditions. Time-lapse imaging allowed us to further identify sub-positions within the G1 and S phases at the time of irradiation by two independent means, and to quantitate the number of MNs by following each cell through M phase until the next G1 phase. Notably, we found that radioresponse was low in late G1 phase, but rapidly increased in early S phase. It then decreased until late S phase and increased in G2 phase. For the first time, we demonstrated the unique fluctuation of radioresponse by the MN assay during the cell cycle in HeLa cells. We discuss the difference between previous clonogenic experiments using M phase-synchronised cell populations and ours, as well as the clinical implications of the present findings.

Isolation and Characterization of Living Cord Blood CD34+ Cells with Telomerase Reverse Transcriptase (TERT) Expression.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 3559-3559
Author(s):  
Marcus Nilsson-Jaras ◽  
Anna Edqvist ◽  
Johan Rebetz ◽  
Bengt Widegren ◽  
Stefan Karlsson ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Progenitor Cells ◽  
Adenoviral Vector ◽  
Telomerase Activity ◽  
Telomerase Reverse Transcriptase ◽  
Cd34 Cells ◽  
M Phase ◽  
Colony Forming Capacity ◽  
Tert Expression ◽  
Vector Transduction

Abstract In the hematopoietic system, telomerase activity is suggested to be differentiation- and proliferation status-dependent. High telomerase activity has been demonstrated in bulk CD34+ progenitor cells. The telomerase activity is mainly controlled at the transcriptional level of the telomerase reverse transcriptase (TERT) gene. In this study, we functionally characterized living cord blood (CB) CD34+ cells with TERT promoter activity by using a TERT reporting adenoviral vector with Ad35 tropism. Such fiber retargeted Ad5F35 vectors allow efficient gene transfer into hematopoietic cells by using the ubiquitously expressed CD46 as a cellular receptor. An adenoviral vector (cTERTdGFP) encoding destabilized EGFP (half-life of 2 hours) under the control of the human TERT promoter and the chicken b-like globin gene insulator was constructed. The background expression of GFP from cTERTdGFP was assessed in the telomerase(−) CB CD15/33+ cells, WI-38 cells and fibroblast cells. Less than 3 percent of these cell types expressed low levels of GFP following transduction with cTERTdGFP in comparison to the control vector Ad5F35-GFP (PGK-promoter) transduced cells (78 to 95% expressed GFP). Under similar conditions, more than 95% of the telomerase(+) A549 cells expressed GFP+ following the cTERTdGFP vector transduction. Therefore, the cTERTdGFP vector allowed d2GFP expression in a telomerase activity-dependent manner. Telomerase activity levels were quantified using the TRAP assay. All transductions were performed at an MOI of 100. The CB CD34+ cells were cultured in serum-free medium supplemented with thrombopoietin. When transduced with the Ad5F35-GFP vector at an MOI of 100, 47±6.7% of the CD34+ cells expressed GFP after two days, while 17±4.3% of the cells expressed GFP following the cTERTdGFP vector transduction. Sorted GFP+ cells following transduction with the cTERTdGFP (TERT sorted) or the Ad5F35-GFP (control sorted) vector were assessed for cell cycle distribution, colony forming capacity and repopulating capacity. Staining for the Ki-67 antigen and 7-AAD revealed that the TERT sorted cells had a greater proportion of cells in the S/G2/M phase of the cell cycle compared to the control sorted cells (55±1.2% versus 37±3.6%, p<0.01), and fewer cells in G0 phase (8.7±2.3% versus 21±3.7%, p<0.05). The colony forming capacity of TERT- and control-sorted cells was similar. Fourteen days following plating of 500 TERT sorted cells, 99±28 BFU-E and 59±18 CFU-G/M colonies were scored compared to the control sorted cells that formed 92±28 BFU-E and 59±11 CFU-G/M colonies. To further assess whether the TERT expressing cells contained repopulating primitive progenitor cells, 1x105 TERT sorted cells were transplanted via tail vein injection into NOD/SCIDBeta2m−/− mice. Human cell reconstitution in the bone marrow was examined at 6 weeks post-transplant in two independent experiments. The TERT sorted cells showed an average of 34±18% (n=9) engraftment with both B- and myeloid-lineage differentiation. Similar engraftment was observed for control sorted cells (35±11%, n=8). In summary, the cTERTdGFP vector allowed isolation of single living primitive hematopoietic progenitor cells with TERT expression. This cell population is enriched for cells in the S/G2/M phase of cell cycle and contains colony-forming progenitor cells and NOD/SCIDBeta2m−/− repopulating progenitor cells.

Decitabine induces cell cycle arrest at the G1 phase via p21WAF1 and the G2/M phase via the p38 MAP kinase pathway

2003 ◽  
Vol 27 (11) ◽  
pp. 999-1007 ◽  
Author(s):  
Donald Lavelle ◽  
Joseph DeSimone ◽  
Maria Hankewych ◽  
Tatiana Kousnetzova ◽  
Yi-Hsiang Chen
Keyword(s):  
Cell Cycle ◽  
Map Kinase ◽  
Cell Cycle Arrest ◽  
P38 Map Kinase ◽  
G1 Phase ◽  
Cycle Arrest ◽  
M Phase ◽  
Map Kinase Pathway ◽  
Kinase Pathway

scholarly journals Estrogen Receptor Beta Displays Cell Cycle-Dependent Expression and Regulates the G1 Phase through a Non-Genomic Mechanism in Prostate Carcinoma Cells

2008 ◽  
Vol 30 (4) ◽  
pp. 349-365 ◽  
Author(s):  
Antoni Hurtado ◽  
Tomàs Pinós ◽  
Anna Barbosa-Desongles ◽  
Sandra López-Avilés ◽  
Jordi Barquinero ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Cell Cycle ◽  
Estrogen Receptor ◽  
Estrogen Receptor Beta ◽  
Mechanisms Of Action ◽  
S Phase ◽  
G1 Phase ◽  
Over Expression ◽  
M Phase ◽  
Receptor Beta

Background: It is well known that estrogens regulate cell cycle progression, but the specific contributions and mechanisms of action of the estrogen receptor beta (ERβ) remain elusive.Methods: We have analyzed the levels of ERβ1 and ERβ2 throughout the cell cycle, as well as the mechanisms of action and the consequences of the over-expression of ERβ1 in the human prostate cancer LNCaP cell line.Results: Both ERβ1 mRNA and protein expression increased from the G1 to the S phase and decreased before entering the G2/M phase, whereas ERβ2 levels decreased during the S phase and increased in the G2/M phase. ERβ1 protein was detected in both the nuclear and non-nuclear fractions, and ERβ2 was found exclusively in the nucleus. Regarding the mechanisms of action, endogenous ERβ was able to activate transcription via ERE during the S phase in a ligand-dependent manner, whereas no changes in AP1 and NFκB transactivation were observed after exposure to estradiol or the specific inhibitor ICI 182,780. Over-expression of either wild type ERβ1 or ERβ1 mutated in the DNA-binding domain caused an arrest in early G1. This arrest was accompanied by the interaction of over-expressed ERβ1 with c-Jun N-terminal protein kinase 1 (JNK1) and a decrease in c-Jun phosphorylation and cyclin D1 expression. The administration of ICI impeded the JNK1–ERβ1 interaction, increased c-Jun phosphorylation and cyclin D1 expression and allowed the cells to progress to late G1, where they became arrested.Conclusions: Our results demonstrate that, in LNCaP prostate cancer cells, both ERβ isoforms are differentially expressed during the cell cycle and that ERβ regulates the G1 phase by a non-genomic mechanism.

scholarly journals Functional Heterogeneity of Human CD34+ Cells Isolated in Subcompartments of the G0 /G1 Phase of the Cell Cycle

Blood ◽  
1997 ◽  
Vol 90 (11) ◽  
pp. 4384-4393 ◽  
Author(s):  
André Gothot ◽  
Robert Pyatt ◽  
Jon McMahel ◽  
Susan Rice ◽  
Edward F. Srour
Keyword(s):  
Cell Cycle ◽  
Ex Vivo ◽  
G1 Phase ◽  
Dna And Rna ◽  
Human Cd34 ◽  
Rna Content ◽  
Cd34 Cells ◽  
Ex Vivo Culture

Using simultaneous Hoechst 33342 (Hst) and Pyronin Y (PY) staining for determination of DNA and RNA content, respectively, human CD34+ cells were isolated in subcompartments of the G0 /G1 phase of the cell cycle by flow cytometric cell sorting. In both bone marrow (BM) and mobilized peripheral blood (MPB) CD34+ cells, primitive long-term hematopoietic culture-initiating cell (LTHC-IC) activity was higher in CD34+ cells isolated in G0 (G0CD34+ cells) than in those residing in G1 (G1CD34+ cells). However, as MPB CD34+ cells displayed a more homogeneous cell-cycle status within the G0 /G1 phase and a relative absence of cells in late G1 , DNA/RNA fractionation was less effective in segregating LTHC-IC in MPB than in BM. BM CD34+ cells belonging to four subcompartments of increasing RNA content within the G0 /G1 phase were evaluated in functional assays. The persistence of CD34 expression in suspension culture was inversely correlated with the initial RNA content of test cells. Multipotential progenitors were present in G0 or early G1 subcompartments, while lineage-restricted granulomonocytic progenitors were more abundant in late G1 . In vitro hematopoiesis was maintained for up to 6 weeks with G0CD34+ cells, whereas production of clonogenic progenitors was more limited in cultures initiated with G1CD34+ cells. To test the hypothesis that primitive LTHC-ICs would reenter a state of relative quiescence after in vitro division, BM CD34+ cells proliferating in ex vivo cultures were identified from their quiescent counterparts by a relative loss of membrane intercalating dye PKH2, and were further fractionated with Hst and PY. The same functional hierarchy was documented within the PKH2dim population whereby LTHC-IC frequency was higher for CD34+ cells reselected in G0 after in vitro division than for CD34+ cells reisolated in G1 or in S/G2 + M. However, the highest LTHC-IC frequency was found in quiescent PKH2bright CD34+ cells. Together, these results support the concept that cells with distinct hematopoietic capabilities follow different pathways during the G0 /G1 phase of the cell cycle both in vivo and during ex vivo culture.

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