The addition of albumin improves Schwann cells viability in nerve cryopreservation

Cell and Tissue Banking volume 19, pages507–517(2018)Cite this article

Abstract

The purpose of the current study was to establish a valid protocol for nerve cryopreservation, and to evaluate if the addition of albumin supposed any advantage in the procedure. We compared a traditional cryopreservation method that uses dimethyl sulfoxide (DMSO) as cryoprotectant, to an alternative method that uses DMSO and albumin. Six Wistar Lewis rats were used to obtain twelve 20 mm fragments of sciatic nerve. In the first group, six fragments were cryopreserved in 199 media with 10% DMSO, with a temperature decreasing rate of 1 °C per minute. In the second group, six fragments were cryopreserved adding 4% human albumin. The unfreezing process consisted of sequential washings with saline in the first group, and saline and 20% albumin in the second group at 37 °C until the crioprotectant was removed. Structural evaluation was performed through histological analysis and electronic microscopy. The viability was assessed with the calcein-AM (CAM) and 4′,6-diamino-2-fenilindol (DAPI) staining. Histological results showed a correct preservation of peripheral nerve architecture and no significant differences were found between the two groups. However, Schwann cells viability showed in the CAM-DAPI staining was significantly superior in the albumin group. The viability of Schwann cells was significantly increased when albumin was added to the nerve cryopreservation protocol. However, no significant structural differences were found between groups. Further studies need to be performed to assess the cryopreserved nerve functionality using this new method.

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Acknowledgements

We would like to thank Alberto Centeno for his assistance and commitment in the excellent care taken of the animals, Catalina Sueiro and Ada Castro for their major support in the assessment of the electronic microscopy images, and Jorge Pombo Otero and Ana Reguero for their invaluable assistance in the interpretation of the histological images.

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Affiliations

  1. Servicio de Cirugía Plástica, Complexo Hospitalario Universitario de A Coruña (CHUAC), Servicio Galego de Saúde (SERGAS), Hospital Universitario de A Coruña, Xubias de Arriba 84, 15006, A Coruña, Spain

    Sara Alicia González Porto, Alba González Rodríguez, Edgar Mauricio Avellaneda Oviedo & Ángel Álvarez Jorge

  2. Biobanco A Coruña- Instituto de Investigación Biomédica de A Coruña (INIBIC), A Coruña, Spain

    Nieves Domenech

  3. Centro de Investigación Biomédica en Red de Enfermedades Cardiovasculares (CIBERCV), Madrid, Spain

    Nieves Domenech

  4. Grupo de Investigación de Proteómica-PBR2-ProteoRed/ISCIII-Servicio de Reumatología, Instituto de Investigación Biomédica de A Coruña (INIBIC), Complexo Hospitalario Universitario de A Coruña (CHUAC), Servicio Galego de Saúde (SERGAS), Universidade da Coruña (UDC), A Coruña, Spain

    Francisco J. Blanco

  5. Grupo de Terapia Celular y Medicina Regenerativa (TCMR-CHUAC), CIBER BBN/ISCIII, Instituto de Investigación Biomédica de A Coruña (INIBIC), Complexo Hospitalario Universitario de A Coruña (CHUAC), Servicio Galego de Saúde (SERGAS), Ciencias Biomédicas, Medicina y Fisioterapia, Facultade de Oza, Universidade da Coruña (UDC), A Coruña, Spain

    María C. Arufe Gonda

  6. Unidad de Criobiología, Banco de Tejidos, Complexo Hospitalario Universitario de A Coruña (CHUAC), Servicio Galego de Saúde (SERGAS), A Coruña, Spain

    Jacinto Sánchez Ibañez & Esther Rendal Vázquez

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  1. Sara Alicia González Porto
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Correspondence to Sara Alicia González Porto.

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González Porto, S.A., Domenech, N., González Rodríguez, A. et al. The addition of albumin improves Schwann cells viability in nerve cryopreservation. Cell Tissue Bank 19, 507–517 (2018). https://doi.org/10.1007/s10561-018-9700-7

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Keywords

  • Cryopreservation
  • Dimethyl sulfoxide
  • Peripheral nerve injuries
  • Serum albumin