title: Isolation of osteoprogenitors from human jaw periosteal cells: a comparison of two magnetic separation methods.
abstract: Human jaw periosteum tissue contains osteoprogenitors that have potential for tissue engineering applications in oral and maxillofacial surgeries. To isolate osteoprogenitor cells from heterogeneous cell populations, we used the specific mesenchymal stem cell antigen-1 (MSCA-1) antibody and compared two magnetic separation methods. We analyzed the obtained MSCA-1(+) and MSCA-1(-) fractions in terms of purity, yield of positive/negative cells and proliferative and mineralization potentials. The analysis of cell viability after separation revealed that the EasySep method yielded higher viability rates, whereas the flow cytometry results showed a higher purity for the MACS-separated cell fractions. The mineralization capacity of the osteogenic induced MSCA-1(+) cells compared with the MSCA-1(-) controls using MACS was 5-fold higher, whereas the same comparison after EasySep showed no significant differences between both fractions. By analyzing cell proliferation, we detected a significant difference between the proliferative potential of the osteogenic cells versus untreated cells after the MACS and EasySep separations. The differentiated cells after MACS separation adjusted their proliferative capacity, whereas the EasySep-separated cells failed to do so. The protein expression analysis showed small differences between the two separation methods. Our findings suggest that MACS is a more suitable separation method to isolate osteoprogenitors from the entire jaw periosteal cell population.
authors.affiliation: Department of Oral and Maxillofacial Surgery, University Hospital of Tübingen, Tübingen, Germany.
authors.forename: Marcus
authors.initials: M
authors.lastname: Olbrich
authors.forename: Melanie
authors.initials: M
authors.lastname: Rieger
authors.forename: Siegmar
authors.initials: S
authors.lastname: Reinert
authors.forename: Dorothea
authors.initials: D
authors.lastname: Alexander
mesh_terms: D000906:Antibodies / Q000737:chemistry; D000954:Antigens, Surface / Q000235:genetics* / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D001706:Biopsy; D002130:Calcium Phosphates / Q000378:metabolism; D002454:Cell Differentiation; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D005434:Flow Cytometry; D015870:Gene Expression; D006801:Humans; D007568:Jaw / Q000166:cytology* / Q000502:physiology; D059346:Magnets; D010521:Periosteum / Q000166:cytology* / Q000502:physiology; D061251:Primary Cell Culture; D013234:Stem Cells / Q000166:cytology* / Q000502:physiology
chemical_list: D000906:Antibodies; D000954:Antigens, Surface; D015415:Biomarkers; D002130:Calcium Phosphates; C540400:W8B2 protein, human