In vitro study of core-shell microcapsules for dental pulp stem cell spheroid formation
Objective:To develop a novel strategy for the construction and cultivation of three-dimensional cell spheroids from dental pulp stem cells(DPSCs)with coaxial-electrospray method.Methods:DPSCs were isolated from clinically collected teeth and then identified through flow cytometry.Core-shell microcapsules were constructed.By adjusting the concentration and flow rates of inter-nal and external phase solutions and cell density,the morphology and size of the microcapsules were observed to get the best prepara-tion condition.Cell viability was detected through live/dead staining and CCK-8 assay,and the cytoskeleton was observed using immu-nofluorescence staining.Results:DPSCs were successfully isolated and cultured.Cell-laden microcapsule with alginate layer and car-boxymethylcellulose core structure was fabricated.When the cell density within microcapsules reached 1×108 cells/mL,the DPSCs within a single microcapsule formed a spheroid spontaneously with a diameter of(101.93±10.11)μm within 24 hours.Live/dead stai-ning showed the cell survival rate didnot decrease within 7 days(P＞0.05).CCK-8 assay revealed an increase in cell proliferation rate to(114.91±7.70)%after 4 days of culture(P＜0.05),which further grew to(169.05±5.29)%at 7 days(P＜0.001).Cells formed dense structures within spheroids.Conclusions:These core-shell microcapsules could promote the formation of DPSCs cell spheroids and maintain their long-term survival.
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