Simple and efficient method for consecutive inactivation–cryopreservation of porcine skin grafts

  • Hiromu Masuoka
  • Naoki MorimotoEmail author
  • Atsushi Mahara
  • Michiharu Sakamoto
  • Toshihito Mitsui
  • Natsuko Kakudo
  • Kenji Kusumoto
  • Tetsuji Yamaoka
Original Article Tissue Engineering / Regenerative Medicine


We previously reported that inactivation treatment by high hydrostatic pressurization (HHP) has potential utility as a novel skin regeneration therapy for various skin tumors. In this study, we evaluated whether glycerol-cryopreservation could be applied in order to preserve inactivated skin by HHP using a porcine model. Twenty full-thickness skin grafts (1.5 × 1.5 cm) were prepared from a minipig. The skin samples were inactivated by the HHP in normal saline or glycerol/fructose solution, followed by cryopreservation for 5 weeks at − 80 °C in each same solution. Another 10 grafts immediately after inactivation were prepared as non-cryopreserved controls. Nine grafts in each group were randomly implanted on the fascia of a host pig and removed at 1, 4 and 11 weeks after grafting. All grafts showed engraftment macroscopically. Hematoxylin eosin staining showed the cellular components in all areas of the dermis at 4 and 11 weeks after grafting, and immunohistochemical staining for CD31 showed the presence of capillaries in the grafts in all groups. The surface and cross-sectional areas of grafts in the normal saline solution cryopreserved group decreased between 1 and 11 weeks, whereas these areas in the glycerol cryopreserved group did not decrease significantly. Glycerol cryopreservation may therefore be a simple and efficient method for preserving porcine skin inactivated by HHP.


High hydrostatic pressure Skin graft Inactivation Preservative fluid Cryopreservation 



This research was supported by a Practical Research for Innovative Cancer Control Grant (17ck0106304h0001) from the Japan Agency for Medical Research and Development (AMED).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interests.


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Copyright information

© The Japanese Society for Artificial Organs 2019

Authors and Affiliations

  1. 1.Department of Plastic and Reconstructive SurgeryKansai Medical UniversityHirakata CityJapan
  2. 2.Department of Plastic and Reconstructive Surgery, Graduate School of MedicineKyoto UniversityKyoto CityJapan
  3. 3.Department of Biomedical EngineeringNational Cerebral and Cardiovascular Center Research InstituteSuita CityJapan

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