A technique for in vitro culture of canine valvular interstitial cells.
OBJECTIVE
To develop a method for in vitro culture of canine valvular interstitial cells (VICs).


METHODS
Canine VICs were isolated from the distal third of the anterior mitral valve leaflet using an explant technique and maintained in cell culture. Molecular phenotyping of the cultured cells was performed using reverse transcription polymerase chain reaction and immunocytochemistry.


RESULTS
Cells resembling fibroblasts migrated from canine mitral valve explants and were maintained in culture for up to eight passages. Establishment of the valve explant required collagen but once established, subsequent passages grew on non-coated plastic plates. At confluence the cultured cells exhibited the characteristic whorled pattern of fibroblasts in culture. The isolated valve cells expressed vimentin but not platelet endothelial cell adhesion molecule or von Willebrand's factor, consistent with the molecular phenotype of VICs.


CONCLUSIONS
VICs can be readily isolated from canine mitral valve leaflets and successfully maintained in culture using standard culture techniques. The described techniques permit the study of bioactive VICs in a controlled environment and may be a useful in vitro model for investigation of cellular and molecular alterations associated with canine chronic degenerative valve disease.
Heaney
Allison M
AM
Department of Clinical Sciences, Kansas State University, College of Veterinary Medicine, Manhattan, KS 66506, USA. aheaney@vetmed.wsu.edu
Bulmer
Barret J
BJ
Ross
Christopher R
CR
Schermerhorn
Thomas
T
D000818:Animals; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods / Q000662:veterinary*; D003094:Collagen; D003470:Culture Media / Q000737:chemistry; D004285:Dogs / Q000502:physiology*; D005347:Fibroblasts; D005786:Gene Expression Regulation / Q000502:physiology; D008943:Mitral Valve / Q000166:cytology*; D014746:Vimentin / Q000235:genetics / Q000378:metabolism
D003470:Culture Media; D014746:Vimentin; D003094:Collagen