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A protocol for umbilical cord tissue cryopreservation as a source of mesenchymal stem cells

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A Correction to this article was published on 19 February 2021

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Abstract

Mesenchymal stem cells (MSC) differentiate into different cell types and have immunomodulatory and paracrine effects. Cryopreservation of umbilical cord tissue as a source of MSC is very promising for regenerative medicine. We aim to evaluate a protocol for cryopreserving this tissue sectioned into small fragments with viable MSC. A total of 723 samples were frozen, thawed and cultured to obtain primary cultures of MSC. These were followed until 90–100% confluence and flow cytometric analysis were performed to confirm the mesenchymal phenotype. Samples in which protocol alterations at the collection of the samples were reported, were excluded for microbial contamination analysis leaving a total of 634 samples composed of 181 vaginal and 453 cesarean births. All cultures reach confluence with a media of 22.57 days and 97% in 28 or fewer days. Evaluated cultures showed low percentage of CD45+ and high of CD73 and CD90. Eight samples were subcultured 4 or 5 times and differentiated to chondrocytes and osteocytes to test differentiation potential with positive results. Umbilical cord tissue collections showed similar microbial profile and risk factors to those reported of umbilical cord blood collections, but with higher contamination frequencies. Cryopreserved tissue samples had viable cells that can be expanded without losing differentiation potential. Higher contamination frequencies compared to umbilical cord blood collection are not surprising, however, microbial load and survival of microorganisms to cryopreservation are expected to be lower.

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Acknowledgments

We are grateful to the staff of Protectia Stem Cells Bank from Cordoba Argentina for sharing their mesenchymal stem cells cryopreservation advances used as a starting point for the optimized protocol presented in this work.

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Authors and Affiliations

  1. Criocenter SRL, Buenos Aires, Argentina

    Diego Raffo & Diego Fernandez Sasso

  2. Centro de Investigaciones Endocrinológicas “Dr. César Bergadá” (CEDIE), CONICET – FEI – División de Endocrinología, Hospital de Niños R. Gutiérrez, Buenos Aires, Argentina

    Diego Raffo

  3. Universidad Abierta Interamericana (UAI), Centro de Altos Estudios en Ciencias Humanas y de la Salud (CAECIHS), Buenos Aires, Argentina

    Andrea Maglioco

  4. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina

    Andrea Maglioco

Authors
  1. Diego Raffo
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  2. Andrea Maglioco
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  3. Diego Fernandez Sasso
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Contributions

R.D, M.A and F.S.D contributed to the design and implementation of the research, analysis of the results and writing of the manuscript. R.D designed and performed the experiments.

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Correspondence to Diego Raffo.

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There are no conflicts of interest to report.

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All procedures performed with human samples were in accordance with the ethical standards of the research committee (CEIC, Comité de Etica en Investigación Clínica, http://www.comitedeeticaceic.com.ar/) and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

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The original online version of this article was revised: The authors names were interchanged in the original article. They should read Diego Raffo, Andrea Maglioco, and Diego Fernandez Sasso. The original article has been corrected.

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Raffo, D., Maglioco, A. & Fernandez Sasso, D. A protocol for umbilical cord tissue cryopreservation as a source of mesenchymal stem cells. Mol Biol Rep 48, 1559–1565 (2021). https://doi.org/10.1007/s11033-020-06079-x

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  • DOI: https://doi.org/10.1007/s11033-020-06079-x

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