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In vitro comparative analysis of cryopreservation of undifferentiated mesenchymal cells derived from human periodontal ligament

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Abstract

Cryopreservation aims to cease all biological functions of living tissues in a reversible and controlled manner, i.e., to permit the recovery of cells by maintaining a high degree of their viability and functional integrity. The objective of this study was to evaluate in vitro the influence of cryopreservation on undifferentiated mesenchymal cells derived from the periodontal ligament of human third molars. Mesenchymal cells were isolated from six healthy teeth and cultured in α-MEM medium supplemented with antibiotics and 15% FBS in a humid atmosphere with 5% CO2 at 37°C. The cells isolated from each tooth were divided into two groups: group I (fresh, non-cryopreserved cells) was immediately cultured, and group II was submitted to cryopreservation for 30 days. The rates of cell adhesion and proliferation were analyzed in the two groups by counting the cells adhered to the wells at 24, 48 and 72 h after plating. The number of cells per well was obtained by counting viable cells in a hemocytometer using the Trypan blue exclusion method. Differences between groups at each time point were evaluated by the Wilcoxon test. The Friedman test was used to determine differences between time points and, if detected, the Wilcoxon test with Bonferroni correction was applied. The results showed no significant difference in the in vitro growth capacity of undifferentiated mesenchymal cells between the two groups. In conclusion, cryopreservation for 30 days had no influence on periodontal ligament mesenchymal cells.

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Acknowledgments

The authors thank BioPol (Laboratório de Biotecnologia de Polímeros Naturais, Departamento de Bioquímica, UFRN) for kindly providing the support on the cell culture equipment.

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Authors and Affiliations

  1. Postgraduate Program in Oral Pathology, Federal University of Rio Grande do Norte, Natal, Rio Grande do Norte, Brazil

    Rodrigo Gadelha Vasconcelos & Marcelo Gadelha Vasconcelos

  2. Postgraduate Program in Dentistry, Federal University of Rio Grande do Norte, Natal, Rio Grande do Norte, Brazil

    Rodrigo Alves Ribeiro

  3. Department of Dentistry, Federal University of Rio Grande do Norte, Natal, Rio Grande do Norte, Brazil

    Kenio Costa Lima

  4. Department of Morphology, Center of Biosciences, Federal University of Rio Grande do Norte, Campus Universitário, Av. Salgado Filho, 3000, Natal/RN, CEP 59072-970, Brazil

    Carlos Augusto Galvão Barboza

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  1. Rodrigo Gadelha Vasconcelos
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  2. Rodrigo Alves Ribeiro
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  3. Marcelo Gadelha Vasconcelos
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  4. Kenio Costa Lima
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  5. Carlos Augusto Galvão Barboza
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Correspondence to Carlos Augusto Galvão Barboza.

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Vasconcelos, R.G., Ribeiro, R.A., Vasconcelos, M.G. et al. In vitro comparative analysis of cryopreservation of undifferentiated mesenchymal cells derived from human periodontal ligament. Cell Tissue Bank 13, 461–469 (2012). https://doi.org/10.1007/s10561-011-9271-3

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  • DOI: https://doi.org/10.1007/s10561-011-9271-3

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