Cryopreservation of the common carotid artery of the rabbit.
We describe experiments on the cryopreservation of the rabbit common carotid artery aimed at improving upon previous results. We describe the design of a double clamp which holds the artery during transportation and storage, preventing twisting, shortening, and collapse of the vessel. The device allowed perfusion with solutions as desired and markedly reduced the extent of endothelial loss during procurement and processing. We also studied the effects of three vehicle solutions; a modified Hanks' solution, a solution originally developed for the cryopreservation of smooth muscle (K-Pipes), and a solution designed for corneal endothelium (CP-Tes). The criteria used to make the assessments were smooth muscle contractility and the structure and function of the vascular endothelium. A new staining method for vascular endothelium (combining propidium iodide with silver nitrate) is described. We found that there was significantly more endothelial cell damage in rabbit carotid arteries frozen in Hanks' solution than in the other solutions, and the recovery of smooth muscle contractility was lowest in the Hanks' group. Arteries cryopreserved using CP-Tes as the vehicle solution showed less endothelial cell damage than arteries preserved with either K-Pipes or Hanks' solution, and these arteries also exhibited the greatest relaxation response to acetylcholine. We conclude that careful handling of the vessels is important; of the solutions studied, CP-Tes is preferred for the cryopreservation of rabbit carotid artery with Me2SO.
Song
Y C
YC
MRC Medical Cryobiology Group, University Department of Surgery, Cambridge, United Kingdom.
Hunt
C J
CJ
Pegg
D E
DE
D000818:Animals; D017536:Carotid Artery, Common / Q000502:physiology / Q000648:ultrastructure; D015925:Cryopreservation / Q000295:instrumentation / Q000379:methods*; D003451:Cryoprotective Agents; D004121:Dimethyl Sulfoxide; D004730:Endothelium, Vascular / Q000502:physiology / Q000648:ultrastructure; D005069:Evaluation Studies as Topic; D005260:Female; D066298:In Vitro Techniques; D007552:Isotonic Solutions; D008297:Male; D008854:Microscopy, Electron; D009119:Muscle Contraction; D009131:Muscle, Smooth, Vascular / Q000502:physiology / Q000648:ultrastructure; D011419:Propidium; D011817:Rabbits; D012835:Silver Nitrate; D012996:Solutions; D013194:Staining and Labeling; D014664:Vasodilation
D003451:Cryoprotective Agents; C072670:Hanks Balanced Salt Solution; D007552:Isotonic Solutions; D012996:Solutions; D011419:Propidium; D012835:Silver Nitrate; D004121:Dimethyl Sulfoxide