scholarly journals 23: Early hematopoietic cells, including megakaryocyte progenitors, are recovered in ALDH bright cell populations isolated by cell sorting from previously frozen umbilical cord blood

2007 ◽  
Vol 13 (2) ◽  
pp. 11
Author(s):  
E. Deibert ◽  
T. Gentry ◽  
S.J. Foster ◽  
J. Kurtzberg ◽  
A.E. Balber
Hematology ◽  
2010 ◽  
Vol 15 (1) ◽  
pp. 11-20 ◽  
Author(s):  
Antonio Alvarado-Moreno ◽  
Antonieta Chávez-González ◽  
Arturo Cérbulo ◽  
Lourdes Arriaga ◽  
Hector Mayani

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 4191-4191
Author(s):  
Bao-An Chen ◽  
Fei Fei ◽  
Cheng-Yin Huang ◽  
Cui-Ping Li ◽  
Xiao-Ping Pei ◽  
...  

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.


2007 ◽  
Vol 16 (2) ◽  
pp. 223-230 ◽  
Author(s):  
Antonio Alvarado-Moreno ◽  
Antonieta Chávez-González ◽  
Arturo Cérbulo ◽  
Lourdes Arriaga ◽  
Hector Mayani

2003 ◽  
Vol 71 (5) ◽  
pp. 341-350 ◽  
Author(s):  
Colin P. McGuckin ◽  
Daniel Pearce ◽  
Nicolas Forraz ◽  
Jennifer A. Tooze ◽  
Suzanne M. Watt ◽  
...  

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 3222-3222
Author(s):  
Ping Mao ◽  
Huaxin Duan ◽  
Caixia Wang ◽  
Tingfen Deng ◽  
Changru Luo ◽  
...  

Abstract Abstract 3222 Poster Board III-159 Previous studies of expansion for hematopoietic stem/progenitor cells showed progenitor cells could reach a great expansion but were mainly performed at small culture volume. The poor cell number of expanded cells even great folds of expansion in a no more than 10 ml culture system limited the application for clinic transplantation. Thereof, we expanded umbilical cord blood (UCB) hematopoietic cells at a large scale in 250 ml bioreactor culture system. The mononuclear cell isolated form fresh UCB samples were ex vivo expanded in 250 ml bioreactor with serum-free medium supplied with SCF, FL3 and TPO at concentration 20 ng/ml. Cell number, surface markers and colony forming potential after 7 days of expansion have been evaluated. The sub-lethally irradiated animal models were used to analyze engraftment capability of HSC by transplant of expanded cells in severe combined immunodeficient and nonobese diabetic (NOD/SCID) mice. The detection of human hematopoietic cells in the bone marrow of the mice was performed at 6 wk after the transplant. We have successfully expanded hematopoietic progenitor cells 5 times using the modified bioreactor. The cell viability showed no obvious variations during culture. Total cells increased from 1.76±0.50 ×108 (range 1.2-2.5) to 4.14±0.83×108 (range 3.7-5.6) after expansion, CD34+ cells from 1.25±0.31 ×106 (range 0.96-1.75) to 3.96±0.78×106 (range 2.88-5.04) and CD133+ cells from 1.55±0.69×106 (range 0.96-2.75) to 4.77±0.88×106 (range 4.00-5.04). The colonies per 105 cells of CFU-E /BFU-E, CFU-GM and CFU-Mix increased from 369.6±71.5 to 1648.5±504.3, 42.8±81.4 to 146.4±54.5, 39.1±10.3 to 144.7±38.8, respectively. The positive expression of CD34+ was 0.7%±0.28% at day 0 and 0.9%±0.34% after culture in bioreactor at day 7 and CD133+ cells was 0.8%±0.24% and 1.1%±0.35%. There was no significant survive rate difference between expanded and nonexpanded cells transplantation group. The analysis of multilineage hematopoiesis showed that transplanted human hematopoietic cells is represented in murine bone marrow cells by detection the percentage of human cells identified with human specific anti CD3/19/33/45/61/71 MoAb by FACS and the human specific gene Alu-1 and Cat-1 by PCR. Disclosures No relevant conflicts of interest to declare.


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