VLA-2 and VLA-5 Cell Adhesion Molecules Expression in CD34+ Cells from Umbilical Cord Blood and from Bone Marrow

2002 ◽  
Vol 20 (2) ◽  
pp. 174-176 ◽  
Author(s):  
C. Pafumi ◽  
R. Mancari ◽  
G. Parisi ◽  
M. Farina ◽  
A. Russo ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Cell Adhesion ◽  
Cord Blood ◽  
Adhesion Molecules ◽  
Umbilical Cord ◽  
Umbilical Cord Blood ◽  
Cell Adhesion Molecules ◽  
Cd34 Cells

scholarly journals Transplantation Induces Profound Changes in the Transcriptional Asset of Hematopoietic Stem Cells: Identification of Specific Signatures Using Machine Learning Techniques

2020 ◽  
Vol 9 (6) ◽  
pp. 1670
Author(s):  
Daniela Cilloni ◽  
Jessica Petiti ◽  
Valentina Campia ◽  
Marina Podestà ◽  
Margherita Squillario ◽  
...  
Keyword(s):  
Machine Learning ◽  
Bone Marrow ◽  
Cord Blood ◽  
Umbilical Cord ◽  
Umbilical Cord Blood ◽  
Machine Learning Techniques ◽  
Specific Gene ◽  
Self Renewal ◽  
Hematopoietic Stem ◽  
Cd34 Cells

During the phase of proliferation needed for hematopoietic reconstitution following transplantation, hematopoietic stem/progenitor cells (HSPC) must express genes involved in stem cell self-renewal. We investigated the expression of genes relevant for self-renewal and expansion of HSPC (operationally defined as CD34+ cells) in steady state and after transplantation. Specifically, we evaluated the expression of ninety-one genes that were analyzed by real-time PCR in CD34+ cells isolated from (i) 12 samples from umbilical cord blood (UCB); (ii) 15 samples from bone marrow healthy donors; (iii) 13 samples from bone marrow after umbilical cord blood transplant (UCBT); and (iv) 29 samples from patients after transplantation with adult hematopoietic cells. The results show that transplanted CD34+ cells from adult cells acquire an asset very different from transplanted CD34+ cells from cord blood. Multivariate machine learning analysis (MMLA) showed that four specific gene signatures can be obtained by comparing the four types of CD34+ cells. In several, but not all cases, transplanted HSPC from UCB overexpress reprogramming genes. However, these remarkable changes do not alter the commitment to hematopoietic lineage. Overall, these results reveal undisclosed aspects of transplantation biology.

scholarly journals Differential effects of recombinant thrombopoietin and bone marrow stromal-conditioned media on neonatal versus adult megakaryocytes

Blood ◽  
2006 ◽  
Vol 108 (10) ◽  
pp. 3360-3362 ◽  
Author(s):  
Karen M. Pastos ◽  
William B. Slayton ◽  
Lisa M. Rimsza ◽  
Linda Young ◽  
Martha C. Sola-Visner
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Cord Blood ◽  
Umbilical Cord ◽  
Umbilical Cord Blood ◽  
Peripheral Blood ◽  
Conditioned Media ◽  
Valuable Source ◽  
Cd34 Cells ◽  
Adult Bone

Abstract Umbilical cord blood (CB) is a valuable source of stem cells for transplantation, but CB transplantations are frequently complicated by delayed platelet engraftment. The reasons underlying this are unclear. We hypothesized that CB- and peripheral-blood (PB)–derived megakaryocytes (MKs) respond differently to the adult hematopoietic microenvironment and to thrombopoietin (Tpo). To test this, we cultured CB- and PB-CD34+ cells in adult bone marrow stromal conditioned media (CM) or unconditioned media (UCM) with increasing concentrations of recombinant Tpo and compared the effects of these conditions on CB-versus PB-MKs. PB-MKs reached highest ploidy in response to UCM + 100 ng/mL rTpo, and the addition of CM inhibited their maturation. In contrast, CB-MKs reached highest ploidy in CM without rTpo, and high rTpo concentrations (> 0.1 ng/mL) inhibited their maturation. This is the first evidence that human neonatal and adult MKs have substantially different biologic responses to Tpo and potentially to other cytokines.

Humanized Sc(Fv)2 Minibody against C-Mpl Successfully Increased Platelet Number in Monkey and Increased Engraftment of Human Umbilical Cord Blood Cells in NOD/SCID Mouse.

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 2322-2322
Author(s):  
Takashi Yoshikubo ◽  
Yoshihiro Matsumoto ◽  
Masahiko Nanami ◽  
Takayuki Sakurai ◽  
Hiroyuki Tsunoda ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Cord Blood ◽  
Progenitor Cells ◽  
Umbilical Cord ◽  
Umbilical Cord Blood ◽  
Platelet Recovery ◽  
Cynomolgus Monkeys ◽  
Cd34 Cells

Abstract Thrombopoietin (TPO, the ligand for c-mpl) is a key factor for megakaryopoiesis. Several clinical trials of TPO have been conducted for thrombocytopenia without much success due to, in part, the production of neutralized antibodies against endogenous TPO, which causes thrombocytopenia. To overcome this problem, we previously demonstrated that mouse type minibody against c-mpl, with an amino acid sequence totally different from TPO, showed megakaryopoiesis and increased platelet numbers in monkey. This time, using CDR grafting, we generated a humanized sc(Fv)2VB22B minibody (huVB22B) against c-mpl for therapeutic use. The new minibody showed almost the same activity in vitro as TPO and the mouse type minibody, confirmed by both a human megakaryocyte cell (CD41+) differentiation assay and a proliferation assay with TPO-dependent cell line, M-07e. Single sc or iv administration of huVB22B to cynomolgus monkeys showed a dose-dependent increase in platelet numbers. Pharmacokinetic analysis showed that the plasma half-life (T1/2) of huVB22B at iv and sc administration to cynomolgus monkeys was 7–8 h and 11–16 h, respectively. After administration of huVB22B, the platelets of these monkeys increased and showed functional aggregation in response to ADP in vitro. Repeated administration of huVB22B (0.2, 2 and 20mg/kg/week) revealed that the increase in platelet level in cynomolgus monkeys was maintained for a month. Very slight reticular fibers in bone marrow were detected in a high dose group (20mg/ kg). No overt changes were detected by toxicity evaluations including clinical pathology and histopathology in 0.2 and 2mg/kg groups. No neutralized activities in plasma were observed during these experiments. Next, we examined the activities of huVB22B on human bone marrow-derived CD34-positive cells (BM-CD34+) and umbilical cord blood-derived CD34-positive cells (UCB-CD34+) in vitro. BM-CD34+ and UCB-CD34+ cells were cultured with huVB22B in serum free medium. HuVB22B induced differentiation of CD41+ cells from BM-CD34+ or UCB-CD34+ cells in a similar dose-dependent manner. However, UCB-CD34+ cells showed greater proliferation in response to huVB22B compared to BM-CD34+ cells. We then examined the in vivo activities of huVB22B on UCB CD34+ cells by treating NOD/SCID mice transplanted with human UCB-CD34+ cells with huVB22B and examining the bone marrow cells of the mice. The results showed that, compared with the control, administration of huVB22B showed an increase in the number of human hematopoietic progenitor cells (CD34+), lymphoid lineage cells (CD19+), and myeloid lineage cells (CD33+) in addition to human CFU-Meg cells (CD41+). These results suggest that c-mpl stimulation in vivo after transplantation might increase engraftment of progenitor cells in the bone marrow microenvironment and subsequently induce differentiation to multilineage cells. Umbilical cord blood transplantation faces frequent complications including a low-level stem/progenitor cell engraftment and delayed platelet recovery. Our results suggest that c-mpl stimulation might be used to increase the engraftment of UCB stem/progenitor cells and shorten the time of platelet recovery following UCB transplantation.

Platelet-Derived Microparticels Stimulate Proliferation of Colony-Forming Unit Granulocyte-Macrophage of Umbilical Cord Blood Hematopoietic Stem Cells.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 4191-4191
Author(s):  
Bao-An Chen ◽  
Fei Fei ◽  
Cheng-Yin Huang ◽  
Cui-Ping Li ◽  
Xiao-Ping Pei ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Cord Blood ◽  
Adhesion Molecules ◽  
Umbilical Cord ◽  
Umbilical Cord Blood ◽  
Hematopoietic Cells ◽  
Control Group ◽  
Hematopoietic Stem ◽  
Hematopoietic Recovery

Abstract Umbilical Cord blood has become an important source of hematopoietic stem-progenitor cells for transplantation, however hematopoietic recovery after transplantation with umbilical cord blood is slower than with bone marrow or mobilized peripheral blood. Adhesion molecules on hematopoietic cells are involved in hematopoietic cells’ homing, which is considered the most important step of hematological recovery. Some articles indicated that expressions of adhesion molecules on CD34+ cells could predict the time to hematopoietic recovery after transplantation with bone marrow and peripheral blood of many adhesion molecules (such as CD62, CXCR4) are significantly lower on umbilical cord blood than on bone marrow. It is a possible reason for the difficulty in hematopoietic recovery after umbilical cord blood transplant. Platelet -derived microparticles (PMPs) are submicroscopic (<1 μm) membrane vesicles released from platelet if they are stimulated with agonists such as thrombin, collagen, or calcium ionophore A23187 or if exposed to high-stress shear forces. PMPs express several platelet-endothelium attachment receptors on their surface, for example, glycoprotein IIb/IIIa (CD41), Ib and IaIIa, and P-selectin (CD62P) and several other platelet relevant receptors such as CXCR4 and PAR-1. Some articles indicate that PMPs can affect the function of hematopoietic stem cells by increasing the adhesion of hematopoietic cells to fibrinogen, which suggests that PMP-transferred CD41 antigen plays an important role in this process. PMPs can also increase the survival of human hematopoietic cells including human CD34+ clonogenic progenitors. In our research, we observe the function of PMP to affect the cloning efficiency of colony-forming unit granulocyte-macrophage (CFU-GM). We adopt different concentrations of Thrombin (2U/ml, 1.5U/ml, 1.0U/ml and 0.5U/ml) to activate the platelet and acquire PMPs. Then PMPs were evaluated by using flow cytometry. Based on the result that stimulation of platelets by Thrombin (1U/ml) can acquire the best efficiency of PMPs, we used this concentration in all subsequent experiments. Umbilical cord mononuclear cells (MNCs) were obtained from healthy donors and isolate the MNCs by Ficoll-Hypaque density gradient centrifugation. Briefly, MNCs incubated with or without PMPs cultured in 2.7% methylcellulose. CFU-GM growth was stimulated with 30% umbilical cord serum, rhIL-3 and rh GM-CSF. Cultures were incubated at 37°C in a fully humidified atmosphere supplemented with 5% CO2. Colonies were counted under an inverted microscope after 7 or 10 days. The research was divided into four groups: 1. control group; 2. PMPs(10μg/ml); 3. PMPs(50μg/ml); 4. PMPs(100μg/ml). The colony formation was enhanced with PMPs and is dependently stimulated with PMPs. The number of colonies in the group of PMPs(100μg/ml) is more than that of other groups. The number of colonies in control group, PMPs(10μg/ml), PMPs(50μg/ml) and PMPs(100μg/ml) are 57.4±3.2, 65.6±5.6, 77.1±1.7 and 87.8±5.0 per 1×105 respectively. These increases in different groups were statistically significant when compared with control group(p<0.05). To sum up, PMPs can affect the cloning efficiency of CFU-GM of umbilical cord hematopoietic stem cells and the efficiencies are depended on the concentration of PMPs.

Nonexpanded primary lung and bone marrow–derived mesenchymal cells promote the engraftment of umbilical cord blood–derived CD34+ cells in NOD/SCID mice

2003 ◽  
Vol 31 (10) ◽  
pp. 881-889 ◽  
Author(s):  
Pieternella S in 't Anker ◽  
Willy A Noort ◽  
Alwine B Kruisselbrink ◽  
Sicco A Scherjon ◽  
Willem Beekhuizen ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Cord Blood ◽  
Umbilical Cord ◽  
Umbilical Cord Blood ◽  
Mesenchymal Cells ◽  
Scid Mice ◽  
Cd34 Cells

The Higher CD34+ Cell Counts in Cord Blood Than Those in Neonatal Peripheral Blood Correlate with Increased Expression of Cytokine Receptors and Cell Adhesion Molecules on the CD34+ Cells in Cord Blood.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4126-4126
Author(s):  
Young-Ho Lee ◽  
Joong-Pyo Kim ◽  
Myung-Hwa Sung ◽  
Ri-Young Ko ◽  
Jin-Yeong Han
Keyword(s):  
Cell Adhesion ◽  
Cord Blood ◽  
Adhesion Molecules ◽  
Peripheral Blood ◽  
Cell Adhesion Molecules ◽  
Mononuclear Cells ◽  
Cytokine Receptors ◽  
Hematopoietic Stem ◽  
Cell Counts ◽  
Cd34 Cells

Abstract The hematologic values of neonatal peripheral blood show a characteristic series of changes during neonatal period. The hematopoietic stem cells in cord blood (CB) are more primitive and abundant than those in neonatal peripheral blood (NB). However, there have been no studies regarding the reasons for those differences. Thus, We investigated the kinetics of nucleated cells and CD34+ cells in CB and neonatal peripheral blood. We obtained the CB and NB samples from 14 normal full-term babies for this study, which were collected immediately after delivery as well as at 24 hours and 48 hours after delivery. We analysed the expressions of CD34, CD34/CXCR4, CD34/CD49d and CD34/CD44 as well as CD3, CD19 and CD16/56 on the isolated mononuclear cells by using FACSort. We also performed CFU-GM counts and apoptotic analysis with isolated mononuclear cells. The total white blood cell counts as well as absolute neutrophil counts of NB at delivery were higher than those of CB (p=0.04) and increased until 24 hours, when decreased until 48 hours after delivery. There were no differences of absolute lymphocyte counts, CD3+ cell and CD19+ cell counts among CB and NB at 0, 24 and 48 hours postnatally, however, the CD16/56+ cell counts were lower in CB than in NB and gradually decreased until 48 hours after delivery (p=0.011). The number of CFU-GM counts (p=0.02) and CD34+ cells (p=0.049) as well as CD34+CXCR4+ cells (p=0.002), CD34+CD49d+ (p=0.001) and CD34+CD44+ cells (p=0.001) in CB were more significantly higher than those of NB at delivery, when decreased gradually until 48 hours after delivery. The incidence of apoptosis were statistically not significant among CB and NB at 0, 24 and 48 hours postnatally. In conclusion, the higher leukocyte counts for several days after delivery were correlated with neutrophilic increment which may be influenced by stressful events during delivery. The more higher CFU-GM counts and CD34+ cell counts in CB than those in NB might be correlated with increased expression of cytokine receptors and cell adhesion molecules on the CD34+ cells in CB.

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