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. 2019 May 7;9:7057. doi: 10.1038/s41598-019-43468-0

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Isolation and characterization of dental pulp cells. (a) Isolation of dental pulp from tooth specimen, (b) colony formation of in vitro cultivated DPCs 72 hours after isolation, (c) confluent layer of DPCs in standard cell culture condition and clearly visible fibroblastic morphology of expanded cells. (d) Surface marker expression of expanded DPCs in vitro (N = 4). The histograms represent individual experiments where surface marker expression of individual donors was quantified by flow cytometry for the positive and negative mesenchymal stromal markers. Cells (P1) were positive for CD105, CD106, CD90, CD146, CD13 and CD44, but negative for CD45, CD14, CD34 and CD31. (e) The percentage of positive cells for the respective surface markers is depicted. The middle left graph shows the gating strategy for alive DPCs. (N = 4 biological replicates). (f) Population doubling was assessed by CFSE assay with measured time points of 0 hours, 24 hours 72 hours and 120 hours. CFSE intensity was lowered due to cell division and thereby a doubling time of 24.99 hours was calculated.