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.
Highlights
Bone grafts used in alveolar and jawbone reconstruction can be generated through tissue engineering approaches [1]
We recently showed that mesenchymal stem cell antigen-1 (MSCA-1)+-enriched human jaw periosteum-derived cells (JPCs) had a higher osteogenic potential compared with MSCA-12 cells [5]
We have previously shown [5] that the MSCA-1+ cell fraction exhibits a higher osteogenic capacity compared with the MSCA-12 subpopulation within the heterogeneous population of JPCs
Summary
Bone grafts used in alveolar and jawbone reconstruction can be generated through tissue engineering approaches [1]. The periosteum contains multipotent mesenchymal stem cells (MSCs) that show the potential to differentiate into different lineages, such as adipogenic, chondrogenic, myogenic and osteogenic tissues [2]. The osteogenic progenitors are restricted to the inner layer, known as the cambium layer, whereas the following zone mainly contains tissue fibroblasts. The third zone consists primarily of collagen fibers [3,4]. A reliable separation of the different layers is difficult due to the low thickness of human jaw periosteum tissue, identifying characteristic markers for the osteogenic progenitors is important to ensure the success of future clinical applications using this stem cell type
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