Feasibility of vitrification as a storage method for tissue-engineered blood vessels.
It is well established that, in multicellular systems, conventional cryopreservation results in damaging ice formation, both in the cells and in the surrounding extracellular matrix. As an alternative to conventional cryopreservation, we performed a feasibility study using vitrification (ice-free cryopreservation) to cryopreserve tissue-engineered blood vessels. Fresh, frozen, and vitrified tissue-engineered blood vessels were compared using histological methods, cellular viability, and mechanical properties. Cryosubstitution methods were used to determine the location of ice in conventionally cryopreserved engineered vessels. Ice formation was negligible (0.0 +/- 0.0% of vessel area) in the vitrified specimens, and extensive (68.3 +/- 4.5% of vessel area) in the extracellular matrix of frozen specimens. The metabolic assay and TUNEL staining results indicated that vitrified tissue had similar viability to fresh controls. The contractility results for vitrified samples were >82.7% of fresh controls and, in marked contrast, the results for frozen samples were only 10.7% of fresh controls (p < 0.001). Passive mechanical testing revealed enhanced tissue strength after both freezing and vitrification. Vitrification is a feasible storage method for tissue-engineered blood vessel constructs, and their successful storage brings these constructs one step closer to clinical utility.
Department of Biomedical Engineering, Duke University, Durham, North Carolina, USA.
Shannon L M
SL
Dahl
Zhenzhen
Z
Chen
Amy K
AK
Solan
Kelvin G M
KG
Brockbank
Laura E
LE
Niklason
Ying C
YC
Song
D000818:Animals; D017209:Apoptosis; D001672:Biocompatible Materials / Q000737:chemistry*; D001696:Biomechanical Phenomena; D002339:Carotid Arteries / Q000166:cytology / Q000502:physiology / Q000648:ultrastructure; D002448:Cell Adhesion; D018929:Cell Culture Techniques; D002470:Cell Survival; D002478:Cells, Cultured; D015925:Cryopreservation / Q000379:methods*; D003470:Culture Media / Q000737:chemistry; D019332:Endothelin-1 / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000166:cytology / Q000502:physiology / Q000648:ultrastructure; D005240:Feasibility Studies; D005615:Freezing; D005947:Glucose / Q000378:metabolism; D009119:Muscle Contraction / Q000187:drug effects; D009131:Muscle, Smooth, Vascular / Q000166:cytology* / Q000502:physiology / Q000648:ultrastructure; D019188:Organ Preservation Solutions; D010208:Papaverine / Q000494:pharmacology; D010539:Permeability; D011100:Polyglycolic Acid / Q000737:chemistry; D013552:Swine; D013997:Time Factors; D023822:Tissue Engineering / Q000379:methods
D001672:Biocompatible Materials; D003470:Culture Media; D019332:Endothelin-1; D019188:Organ Preservation Solutions; D011100:Polyglycolic Acid; D010208:Papaverine; D005947:Glucose