Separation of mesenchymal stem cells with magnetic nanosorbents carrying CD105 and CD73 antibodies in flow-through and batch systems.
The aim of this study is to develop magnetically loaded nanosorbents carrying specific monoclonal antibodies (namely CD105 and CD73) for separation of mesenchymal stem cells from cell suspensions. Super-paramagnetic magnetite (Fe3O4) nanoparticles were produced and then coated with a polymer layer containing carboxylic acid functional groups (average diameter: 153 nm and polydispersity index: 0.229). In order to obtain the nanosorbents, the monoclonal antibodies were immobilized via these functional groups with quite high coupling efficiencies up to 80%. These nanosorbents and also a commercially available one (i.e., microbeads carrying CD105 antibodies from Miltenyi Biotec., Germany) were used for separation of CD105+ and CD73+ mesenchymal stem cells from model cell suspension composed of peripheral blood (97.6%), human bone marrow cells (1.2%) and fibroblastic cells (1.2%). The initial concentrations of the CD105+ and CD73+ cells in this suspension were measured as 5.86% and 6.56%, respectively. A flow-through separation system and a very simple homemade batch separator unit were used. We were able to increase the concentration of CD105+ cells up to about 86% in the flow-through separation system with the nanosorbents produced in this study, which was even significantly better than the commercial one. The separation efficiencies were also very high, especially for the CD73+ cells (reached to about 64%) with the very simple and inexpensive homemade batch unit.
Odabaş
S
S
Hacettepe University, Department of Chemical Engineering and Bioengineering Division and Biyomedtek, Ankara, Turkey.
Sayar
F
F
Güven
G
G
Yanikkaya-Demirel
G
G
Pişkin
E
E
D015720:5'-Nucleotidase / Q000276:immunology*; D000906:Antibodies / Q000737:chemistry / Q000276:immunology*; D015703:Antigens, CD / Q000276:immunology*; D002469:Cell Separation / Q000295:instrumentation / Q000379:methods*; D000071063:Endoglin; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000276:immunology; D018448:Models, Immunological; D036103:Nanotechnology / Q000295:instrumentation / Q000379:methods; D011956:Receptors, Cell Surface / Q000276:immunology*; D015203:Reproducibility of Results
D000906:Antibodies; D015703:Antigens, CD; C064206:ENG protein, human; D000071063:Endoglin; D011956:Receptors, Cell Surface; D015720:5'-Nucleotidase