Application of Cryopreservation Technique in the Preservation of Rat Limbs

Abstract

Objective: This study aims to observe the physiological and pathological changes of severed fingers (limbs) under different storage conditions through animal experiments, and to screen out the best preservation conditions. Medthods: Sixty healthy adult male Sprague-Dawley rats were selected and evenly divided into 4 preservation groups, including conventional low-temperature dry (CLTD), the university of wisconsin (UW) solution, cryopreservation and cryopreservation + UW solution preservation group. After harvesting the limbs, were preservated for 72 h and 7 days, respectively. Then the limbs were thawed and replanted in situ. Sciatic nerves were collected for hematoxylin and eosin (HE) staining, and observed the changes in tissue morphology. Results: Replantation was successful in 11 out of 15 rats (73%) in cryopreservation + UW group, and the walking function of the 9 (82%) rats in cryopreservation + UW group were significantly better than that of the cryopreservation preservation group. In addition, the HE staining results shown that the CLTD group nerve bundles were morphologically damaged, and there were more acellular structures and tissue fragments; the UW group nerve bundles were less injured and the perineurium was more complete and orderly; The nerve bundles in the cryopreservation group and cryopreservation + UW group are tightly arranged and the tissue morphology is regular; Compared with the cryopreservation + UW group, the complete of the cryopreservation group is not well. Conclusions: The cryopreservation technology combined with UW solution is a new and effective method for the severed limbs preserving.