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.

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.

Expansion and Multipotencial Differentiation of Mesenchymal Stem Cells Isolated from Patients after High Dose Chemotherapy.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 4259-4259
Author(s):  
Karen L. Prata ◽  
Maristela D. Orellana ◽  
Aparecida M. Fontes ◽  
Karina R. Solano ◽  
Simone Kashima ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
High Dose Chemotherapy ◽  
High Dose ◽  
Cd34 Cells ◽  
Significant Difference ◽  
Cell Cycle Status ◽  
Supportive Function

Abstract Background. High dose chemotherapy (HDCT) followed by autologous PBSC rescue has been increasingly used for the treatment of several human diseases. However, little is known on the extent of this therapy on the marrow mesenchymal stem cells (MSCs). Aims. To evaluate the feasibility of expansion and multipotencial differentiation of MSCs isolated from patients after HDCT. Patients and Methods. Twelve lymphoma’s patients (LP) free of disease in bone marrow (BM) were enrolled in the study. They were submitted to BEAM’s protocol with autologous PBSC rescue 28 to 1836 days before the sample collection. Six normal bone marrow donors (ND) were used as controls. The LP and ND median age were 37.5 (range 22–49) and 31.5 years old (range 23–42), respectively. MSCs were isolated by plastic adherence and expanded ex vivo by cultivation in flasks with α-MEM with 15% fetal bovine serum. Media was changed every 3–4 days. At 90% confluence, the cells were re-plated and expanded. The isolation efficiency, colony-forming unit-fibroblast (CFU-F) frequency, growth kinetics, phenotypic characteristics, cell cycle status, multi-lineage differentiation capacity as well as hematopoiesis-supportive function were determined and compared with those of ND-MSCs. This study protocol and the consent form were approved by the institution ethics committees. Results. The results were analyzed by Mann-Whitney test and are expressed as median (range) to LP and ND, respectively. MSCs were successful isolated from all BM samples collected for this study. The cell population showed typical fibroblast-like morphology, appearing as an adherent, spindle shaped cell layer and growing to confluence after a few weeks of culture. The number of CFU-F found at 14 days of culture were 0.94 (0.00–3.75) and 1.25 (0.13–9.25) x10−5 nucleated cells (p = 0.4421). The doubling time between the 1st and 2nd passages was 80.66 (34.08–195.35) and 46.30 (36.36–270.59) hours (p = 0.1025). The cell clones proliferated extensively until 8.17 (1.81–28.27) and 18.11 (11.85–27.48) population doublings (p = 0.0668) in 71.50 (46–88) and 81 (57–103) cultivation days (p = 0.1505). Immunophenotypically, these cells were positive for the CD73, CD105, CD90, CD29, CD13, CD44, CD49e, CD54, HLA-class 1 and Stro-1 markers and negative for CD34, CD45, CD14, CD51/61, HLA-DR and KDR. Regarding the cell cycle status, 85.63 (63.19–92.17) and 82.41 (82.19–87.02) % were in GO-G1 phase (p = 1,000), while only 12.17 (3.33–36.81) and 10.67 (6.59–12.05) % were in S phase (p = 0,6828). All samples tested were capable of differentiating along adipogenic, osteogenic and chondrogenic lineages in vitro, demonstrated by morphology, cyto- and imunohistochemistry or RT-PCR reaction (PPARg and osteopontin genes expression). After co-culture with CD34+ cord blood cells for 1 and 4 weeks, no significant difference CD34+ expansion or colony-forming cells (BFU-E or CFU-GM) were observed between the CD34+ cells/LP-MSCs and CD34+ cells/ND-MSCs co-cultures with cytokines or not. Interpretation and Conclusions. Our results demonstrate that is possible to cultivate and expand MSCs with multipotential differentiation capabilities and hematopoiesis-supportive function from patients after HDCT. Despite there were no significant differences in the median values between LP and ND, the comparative study indicates a possible damage in MSCs by HDCT.

scholarly journals Cell cycle status of CD34+ cells in human fetal bone marrow

2001 ◽  
Vol 65 (2) ◽  
pp. 159-163 ◽  
Author(s):  
Joyce M Koenig ◽  
Benjamin Luttge ◽  
Neal A Benson ◽  
Robert D Christensen
Keyword(s):  
Cell Cycle ◽  
Bone Marrow ◽  
Fetal Bone ◽  
Cd34 Cells ◽  
Cell Cycle Status

Differential Maintenance of Cell Cycle Status of Human Bone Marrow- and Umbilical Cord Blood-Derived CD34+ Cells after Homing to the Bone Marrow Microenvironment.

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 1327-1327
Author(s):  
Bradley A. Poteat ◽  
Brahmananda Reddy Chitteti ◽  
Karen E. Pollok ◽  
Attaya Suvannasankha ◽  
Edward F. Srour
Keyword(s):  
Cell Cycle ◽  
Bone Marrow ◽  
Cord Blood ◽  
Umbilical Cord Blood ◽  
Cell Cycle Progression ◽  
Cycle Progression ◽  
Cd34 Cells ◽  
Cell Cycle Status ◽  
Cells Cultured

Abstract Transplanted human bone marrow (BM) CD34+ cells remain mitotically quiescent up to 72h following their homing to the BM of irradiated NOD/SCID mice (Blood2002;99(5):1585). To investigate whether umbilical cord blood (UCB)-derived CD34+ cells behave similarly and to assess the impact of the BM microenvironment on this observed temporary cell cycle arrest, we examined the cell cycle status of BM- and UCB-derived CD34+ cells recovered from the BM of irradiated (325 cGy) and non-irradiated NOD/SCID IL2Rγnull (NS2) mice 20h after transplantation (AT). To understand the molecular control of this sustained, but short-lived induced quiescence, expression of cell cycle-related proteins among BM-homed cells was examined by real-time quantitative PCR. Freshly isolated CD34+ cells were stained with CFSE-1 and transplanted into normal or conditioned NS2 mice or cultured in vitro with and without SCF, Flt3, GM-CSF, and IL-3. BM was harvested from NS2 recipients 20h AT and CFSE+ cells were recovered by cell sorting. Cell cycle status was assessed by PI staining in a fraction of recovered CFSE+ and cells cultured in vitro and mRNA was isolated from the remaining cells in both groups. The proportion of cells in G0/G1 phase of cell cycle among BM CD34+ cells was maintained in recipient BM 20h AT regardless of whether the microenvironment was irradiated or not (fresh: 83.7% ± 4.4%, n = 14; irradiated: 82.7% ± 5.3%, n = 6; non-irradiated: 84.4% ± 4.1%, n = 10), suggesting that exit of BM-homed cells from G0/G1 is either a strictly cell intrinsic property or is a phenomenon mediated by microenvironmental modulators present in both intact and injured BM. However, cultured BM CD34+ cells cycled efficiently such that after 20h, only 75.5% ± 5.7%, n = 12 remained in G0/G1. Surprisingly, BM-homed UCB CD34+ cells did not remain quiescent and at 20h AT, only 84.85% ± 13.1% (n = 5) were in G0/G1 compared to 97.0% ± 1.8% (n = 14) for freshly isolated cells, suggesting that different cell cycle regulatory mechanisms control BM versus UCB CD34+ cells. UCB CD34+ cells recovered from the BM of irradiated recipient mice 20h AT had increased levels of Bcl-2 and CDKN1B (p27) while those of CDKN1C (p57) and p53 were decreased relative to those isolated from non-irradiated recipients. These findings suggest that at least in the case of UCB CD34+ cells, different cell cycle regulatory networks may impact cell cycle progression of these cells in an irradiated versus intact BM microenvironment. Compared to freshly isolated BM CD34+ cells, those recovered from the marrow of non-irradiated recipients 20h AT had significantly elevated levels of mRNA for CDKN1a (p21), CDKN1B (p27), p53, and N-cadherin, while the mRNA levels for these molecules in comparable cells cultured in vitro for 20h was relatively unchanged. Taken together, these data illustrate that human CD34+ cells from different tissues behave differently during the first few hours following their homing to the BM and that this behavior may be partially regulated by the status of the microenvironment. Furthermore, data from BM-derived CD34+ cells suggest that following homing, active repression of cell cycle progression may be mediated by induced upregulation of p21 and p27. Mechanisms leading to the upregulation of these cell cycle regulatory molecules remain to be investigated.

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 Revisiting Old Practices: More Restricted Indication of Preoperative Autologous Blood Donation in Healthy Bone Marrow Donors According to Baseline Hemoglobin Levels

Blood ◽  
2018 ◽  
Vol 132 (Supplement 1) ◽  
pp. 3819-3819
Author(s):  
Isadora Olenscki Gilli ◽  
Afonso Celso Vigorito ◽  
Bruno Deltreggia Benites
Keyword(s):  
Bone Marrow ◽  
Blood Donation ◽  
Conflicts Of Interest ◽  
Autologous Blood ◽  
Allogeneic Blood ◽  
Autologous Blood Donation ◽  
Roc Curve Analysis ◽  
Preoperative Autologous Blood Donation ◽  
Allogeneic Transfusion ◽  
Cutoff Points

Abstract Background: Despite the cost-effectiveness of preoperative autologous blood donation (PAD) having been intensely debated over the last decade (due to a significant reduction in the risk of transfusion-transmitted diseases), no consensus has been reached as to the risk-benefit status of PAD for healthy bone marrow donors. There is concern regarding the occurrence of pre-procedure anemia caused by the autologous donation itself, consequently increasing the risk of transfusion, and the possibility of unnecessary hospital costs in the case of donors with collected and non-transfused red blood cell concentrates (RBC). Thus, this study aimed to evaluate the changes in hemoglobin levels related to PAD in bone marrow donors followed at our institution and to seek a hemoglobin cutoff with a predictive power for the actual need for this procedure. Study Design and Methods: We conducted a retrospective study at the Hematology and Transfusion Medicine Center at the University of Campinas, Brazil, evaluating the data for all bone marrow donors registered at our institution between 2002 and 2016 who had donated at least one unit of autologous blood. Mean Hgb values were compared separately for donors who donated 1 or 2 units, at 3 time-points: before PAD collection, the morning before marrow harvest and soon after harvest. ROC curves were used to investigate possible Hgb cutoff points for prediction of transfusion requirement. Results: Donors identified in the study period comprised 80 individuals: 76 collected only 1 unit and 4 individuals collected 2 autologous units. Mean baseline Hgb values did not differ significantly between the 2 groups [1 unit: 14.9(12.3-18.1) x 2 units: 13.57 (12.6-14.8), p=0.069]. After PAD collection, there was a significant drop in Hgb levels for the whole cohort of donors [14.82 (12-18.1) x 12.75 (8.9-17.4), p<0.001], more pronounced for the group that donated 2 units (Hgb levels at this time point, 1 unit: 12.8(8.9-17.4) x 2 units: 11.55(11.2-12.1), p=0.045). However, after marrow harvest, Hgb levels were similar for the 2 groups, and despite significant declines in Hgb levels, none of the donors in our cohort required allogeneic transfusion and only 61.2% required autologous transfusion. To further evaluate the use of Hgb levels as a predictor for the efficacy of collecting autologous blood, ROC curve analysis identified baseline Hgb< 14.35g/dL as the sensitive cutoff to predict the need for transfusion after marrow harvest. These levels had a sensitivity of 52% and specificity of 80.4%, p=0.001 and OR=4.38 (95% confidence interval: 1.65-12.59). Conclusion: In the current era of Patient Blood Management, mechanisms that prevent allogeneic blood transfusions, such as PAD, must be explored and debated, since they reduce patient exposure to risks related to allogeneic blood. Our analysis demonstrates the possibility of using hemoglobin thresholds as cutoff points for indication of PAD, tending to a more cost-effective approach. Furthermore, despite significant declines in Hgb levels after PAD, none of the donors in our cohort required allogeneic transfusion, demonstrating the safety of this procedure. Thus, the indication of PAD remains an option for those donors who feel insecure despite higher baseline Hgb levels. Disclosures No relevant conflicts of interest to declare.

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