Cell viability mapping within long-term heart valve organ cultures.
OBJECTIVE
Organ cultures maintain cells within their native microstructural environment, and thus offer greater potential for studying tissue disease and remodeling than do monolayer cell cultures or pathological examinations of diseased tissue. To validate an in-vitro heart valve organ culture model, cell viability was examined within valve tissues over sustained culture periods.


METHODS
Following culture of blocks of valve tissue for 1 to 49 days, cross-sections were cut with a vibratome, stained with a LIVE/DEAD kit, and imaged with confocal microscopy to quantify the number of live and dead cells present.


RESULTS
In numerous organ cultures, valvular interstitial cells were found to be viable beyond 30 days. Live cells were abundant in the central region of the valve, but more sparse in the deepest central regions. Dead cells were found mainly on the surface of both fresh tissues and tissues after prolonged culture, with few dead cells occurring centrally.


CONCLUSIONS
This is the first reported mapping of cell viability within heart valve organ cultures, and results suggest that extended organ culture of valve leaflets is indeed possible. The derived viability staining methods have wide applicability for organ cultures of other tissues as well as tissue-engineered matrices.
Allison
David D
DD
Department of Biomedical Engineering, The University of Iowa, Iowa City, USA.
Drazba
Judith A
JA
Vesely
Ivan
I
Kader
Khalid N
KN
Grande-Allen
K Jane
KJ
D000818:Animals; D001021:Aortic Valve / Q000166:cytology* / Q000378:metabolism; D002417:Cattle; D002452:Cell Count; D002470:Cell Survival / Q000502:physiology; D002478:Cells, Cultured; D004396:Coloring Agents; D004285:Dogs; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism; D004801:Eosine Yellowish-(YS); D006416:Hematoxylin; D006801:Humans; D008855:Microscopy, Electron, Scanning; D008943:Mitral Valve / Q000166:cytology* / Q000378:metabolism; D008955:Models, Cardiovascular; D009924:Organ Culture Techniques; D013552:Swine; D013995:Time; D014261:Tricuspid Valve / Q000166:cytology* / Q000378:metabolism
D004396:Coloring Agents; D004801:Eosine Yellowish-(YS); D006416:Hematoxylin