Cryopreservation of composite tissue transplants

Brian Rinker, Xiangdong D. Cui, Michael L. Cibull, Betsy F. Fink, Dayong Y. Gao, Henry C. Vasconez

Research output: Contribution to journalArticlepeer-review

13 Scopus citations


Composite tissue allotransplantation holds great promise for upper extremity reconstruction but is limited by donor part availability. Cryopreservation may increase the availability of donor parts and even reduce antigenicity. The purpose of the study was to evaluate the viability of cryopreserved composite tissues and to demonstrate the feasibility of microvascular isotransplantation of cryopreserved composite flaps. Twenty epigastric flaps were harvested from Lewis rats. Ten flaps were analyzed fresh. Ten flaps were perfused with dimethyl sulfoxide (DMSO)/trehelose cryoprotectant agent (CPA), frozen by controlled cooling to -140°C, and stored for 2 weeks. Flaps were evaluated by factor VIII endothelial staining and MTT tetrazolium salt assay. For the in vivo phase, 30 flaps were harvested. Ten were transplanted fresh to isogenetic recipient animals, ten were perfused with CPA and transplanted, and ten were cryopreserved for 2 weeks, thawed, and transplanted. All cryopreserved samples displayed intact vascular endothelia on factor VIII staining. On MTT analysis, the epithelial viability index for the cryopreserved samples was not significantly different from fresh controls (p = 0.12). All freshly transplanted flaps (10/10) were viable at 60 days. Nine of ten flaps in the perfused/transplanted group were viable at 60 days. Survival of cryopreserved/transplanted flaps ranged from 5 to 60 days. The skin and vascular endothelial components of composite tissue flaps appear to retain their viability after cryopreservation. The in vivo studies demonstrate that the long-term survival of cryopreserved composite tissue transplants is feasible and support an indirect injury, rather than direct injury from freezing or cryoprotectant agents, as the mechanism of flap loss.

Original languageEnglish
Pages (from-to)17-23
Number of pages7
Issue number1
StatePublished - Mar 2008

Bibliographical note

Funding Information:
Acknowledgement This work is supported by an American Society for Surgery of the Hand Basic Science Research Grant.


  • Composite tissue transplantation
  • Cryopreservation
  • Epigastric flap

ASJC Scopus subject areas

  • Surgery
  • Orthopedics and Sports Medicine


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