The addition of albumin improves Schwann cells viability in nerve cryopreservation
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The addition of albumin improves Schwann cells viability in nerve cryopreservation Sara Alicia Gonza´lez Porto . Nieves Domenech . Alba Gonza´lez Rodrı´guez . Edgar Mauricio Avellaneda Oviedo . Francisco J. Blanco . Marı´a C. Arufe Gonda . ´ ngel A ´ lvarez Jorge . Jacinto Sa´nchez Iban˜ez . Esther Rendal Va´zquez A
Received: 6 November 2017 / Accepted: 21 April 2018 Ó Springer Science+Business Media B.V., part of Springer Nature 2018
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 40 ,6-diamino-2-
S. A. Gonza´lez Porto (&) A. Gonza´lez Rodrı´guez ´. A ´ lvarez Jorge E. M. Avellaneda Oviedo A Servicio de Cirugı´a Pla´stica, Complexo Hospitalario Universitario de A Corun˜a (CHUAC), Servicio Galego de Sau´de (SERGAS), Hospital Universitario de A Corun˜a, Xubias de Arriba 84, 15006 A Corun˜a, Spain e-mail: [email protected]; [email protected]
Complexo Hospitalario Universitario de A Corun˜a (CHUAC), Servicio Galego de Sau´de (SERGAS), Universidade da Corun˜a (UDC), A Corun˜a, Spain
N. Domenech Biobanco A Corun˜a- Instituto de Investigacio´n Biome´dica de A Corun˜a (INIBIC), A Corun˜a, Spain
M. C. Arufe Gonda Grupo de Terapia Celular y Medicina Regenerativa (TCMR-CHUAC), CIBER BBN/ISCIII, Instituto de Investigacio´n Biome´dica de A Corun˜a (INIBIC), Complexo Hospitalario Universitario de A Corun˜a (CHUAC), Servicio Galego de Sau´de (SERGAS), Ciencias Biome´dicas, Medicina y Fisioterapia, Facultade de Oza, Universidade da Corun˜a (UDC), A Corun˜a, Spain
N. Domenech Centro de Investigacio´n Biome´dica en Red de Enfermedades Cardiovasculares (CIBERCV), Madrid, Spain
J. Sa´nchez Iban˜ez E. Rendal Va´zquez Unidad de Criobiologı´a, Banco de Tejidos, Complexo Hospitalario Universitario de A Corun˜a (CHUAC), Servicio Galego de Sau´de (SERGAS), A Corun˜a, Spain
F. J. Blanco Grupo de Investigacio´n de Proteo´mica-PBR2-ProteoRed/ ISCIII-Servicio de Reumatologı´a, Instituto de Investigacio´n Biome´dica de A Corun˜a (INIBIC),
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fenilindol (DAPI) staining. Histological results showed a correct preservation of peripheral nerve architecture and no significant differenc
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