Date of Award

Fall 1992

Document Type

Thesis

Degree Name

Master of Science (MS)

Department

Biological Sciences

Program/Concentration

Biology

Committee Director

Lioyd Wolfinbarger, Jr.

Committee Member

John D. Coulson

Committee Member

Ann E. Campbell

Committee Member

Keith A. Carson

Call Number for Print

Special Collections LD4331.B46 G362

Abstract

Segments of bovine calf aortas and pulmonary arteries were cryopreserved at -196° C for two weeks to 18 months and subsequently thawed. Cellular morphology, viability, and growth potential in these segments were then compared with the same attributes in segments from fresh (control) arteries. Scanning electron microscopy revealed no disruption of intimal surfaces attributable to cryopreservation. Endothelial cells were successfully cultured from all cryopreserved arterial segments. Endothelial monolayers grown from cryopreserved arteries had a cobblestone appearance and expressed Factor VIII antigen but not smooth muscle alpha-actin. Plating efficiency in primary cultures of endothelial cells was lower for cryopreserved arteries than fresh arteries. However, endothelial cells from cryopreserved arteries proliferated more rapidly than and achieved final numbers similar to cells from fresh arteries. Cryopreserved medial explants yielded proliferating smooth muscle cells less frequently than fresh medial explants. Smooth muscle cells from cryopreserved arteries proliferated more slowly and to lower final densities than cells from fresh arteries. Pulmonary artery fibroblasts proliferated less frequently from cryopreserved adventitial explants. However, aortic fibroblasts proliferated as frequently from cryopreserved as from fresh explants. Cryopreserved fibroblasts from both types of arteries proliferated more slowly and to lower final densities than cells from fresh arteries. Smooth muscle cells and fibroblasts from fresh and cryopreserved arteries appeared indistinguishable in culture, and both cell types expressed smooth muscle alpha-actin. Smooth muscle cells were negative for Factor VIII antigen. These observations indicate persistent albeit decreased viability and growth potential in the three layers of the cryopreserved bovine arterial wall.

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DOI

10.25777/tadc-6778

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