ABSTRACT
Objective:To investigate the effect of different collection procedures of COBE Spectra blood cell separator on collection of autologous peripheral blood hematopoietic stem cells in children.Methods:The clinical data of 10 children who were collected autologous peripheral blood hematopoietic stem cells with the monocytes (MNC) or fully automatic peripheral blood stem cells (AutoPBSC) programs of COBE Spectra blood cell separator in the Children's Hospital of Soochow University from July 2018 to January 2020 were retrospectively analyzed, and the children were 3-10 years old with the body weight of 15-31kg. There were 5 cases in MNC group and 5 cases in AutoPBSC group.Results:Autologous peripheral blood hematopoietic stem cells were collected for 25 times, with an average of 2.5 times (1-4 times), 10 times in MNC group and 15 times in AutoPBSC group. The number of stem cells [the median (the range)] before collection was 19.3/μl (3.5-129.0/μl) in MNC group and 9.4/μl (2.2-38.7/μl) in AutoPBSC group. The number of CD34 + cells of single collection was 1.22×10 6/kg (0.18×10 6/kg-6.30×10 6/kg) in MCN group and 0.85×10 6/kg (0.13×10 6/kg-2.64×10 6/kg) in AutoPBSC group. Correlation analysis showed that there was a positive correlation between the number of collected CD34 + cells and the number of stem cells before collection (AutoPBSC group: r=0.921, P < 0.01; MNC group: r=0.833, P=0.003). The collection efficiency was 5.4% (3.4%-11.2%) in MNC group and 10.4% (4.7%-13.9%) in AutoPBSC group, and the difference was statistically significant ( Z=2.163, P = 0.031). Conclusion:The collection effect of children's autologous peripheral blood hematopoietic stem cells with COBE Spectra blood cell separator AutoPBSC program is better than that with MNC program.
ABSTRACT
The ability to generate dendritic cells (DCs) in sizeable numbers has enormous implications for the development of clinically-effective antigen presentation procedures for cancer immunotherapy. We evaluated the generation of immunostimulatory DCs from peripheral blood CD34+ cells collected from healthy donors. CD34+ cells purified from leukapheresis product were seeded at 1 x 10(4) cells/mL in complete medium supplemented with GM-CSF, TNF alpha, IL-4, c-kit ligand, and flt3 ligand (FL). By day 14 of culture in the presence of GM-CSF + TNF alpha, the total cell number increased by 23.4 +/- 5.4-fold compared to the starting number of CD34+ cells. When the c-kit and FL were added to GM-CSF and TNF alpha, the cell number increased by 109.8 +/- 11.2-fold without affecting the immunophenotype of recovered cells. Flow cytometric analysis indicated that cells with the markers of mature dendritic cells, i.e., CD1a +CD14 -HLA-DR+, and CD80+CD86+HLA-DR+, constituted 49.0% +/- 7.5%, and 38.9% +/- 6.5%, respectively. This pattern of expression of surface antigen was unchanged whether the c-kit ligand and/or FL was added. The irradiated CD1a+HLA-DR+ cells recovered from in vitro cultures elicit a vigorous proliferation of allogeneic peripheral blood T-cells, irrespective of cytokine combinations. These findings provide advantageous tools for the large-scale generation of DCs that are potentially usable for clinical protocols of immunotherapy or vaccination in patients undergoing cancer treatment.