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Seeding of Biosynthetic Heart Valves with Endothelial Cells Derived From Sheep Circulating Mononuclear Cells
Abstract   Peer reviewed

Seeding of Biosynthetic Heart Valves with Endothelial Cells Derived From Sheep Circulating Mononuclear Cells

M Passmore, M Nataatmadja, Yoke Lin Fung, B Garlick and J F Fraser
Heart, Lung and Circulation, Vol.20(12), pp.780-781
Australasian Society of Cardiac and Thoracic Surgeons (ASCTS) Annual Scientific Meeting, 2011 (Queenstown, New Zealand, 07-Aug-2011–10-Aug-2011)
2011
DOI: https://doi.org/10.1016/j.hlc.2011.08.019
url
https://doi.org/10.1016/j.hlc.2011.08.019View
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Abstract

Cardiovascular Medicine and Haematology Public Health and Health Services
Introduction: The use of bioprosthetic valves avoids the necessity of warfarinisation, but they often stenose more rapidly, necessitating re-operation. Absence of an endothelial lining increases the risk of early prosthetic valve endocarditis and valve calcification and degeneration. This study aims to isolate and culture circulating sheep mononuclear cells and differentiate them into endothelial cells before seeding them onto the surface of bioprosthetic heart valves to improve their performance. Method: Mononuclear cells were extracted from sheep peripheral blood and cultured on plates coated with sheep Myogel (a muscle derivedmatrix from the BernardO'Brien Institute, Melbourne) to improve cell attachment. They were cultured in endothelial growth media for 21 days to encourage endothelial cell differentiation and then seeded directly onto pieces of mitral valve (n = 10) coated with either Myogel or Poly-L-lysine, for 28 days. They were subsequently fixed in formalin, embedded in paraffin and stained with haematoxylin and eosin. Results: Circulating mononuclear cells were successfully cultured from sheep peripheral blood. Patches of cells with the characteristic cobblestone appearance of endothelial cells were seen after 21 days of culture in endothelial growth media. Haematoxylin and eosin staining of the seeded Poly-L-lysine coated valve showed cells lining the length of the valve. There were no cells lining the Myogel coated valve. Discussion: We have been able to successfully culture mononuclear cells from sheep peripheral blood and differentiate them into endothelial cells. Sections of the Poly-L-lysine coated mitral valve were also successfully seeded with cultured endothelial cells. No cells however were detected lining the Myogel coated valve. As the surface area included for analysiswas limited by the number of cut sections, it is possible that areas of cultured epithelial cells on the Myogel coated valve surface may have been missed. Scanning electron microscopy could be employed to visualise the whole surface of the valve to obtain a clearer idea of cell attachment. Cell retention could also be improved by dynamic seeding where valves are regularly inverted and by employing multiple rounds of seeding. Future studies will need to establish the phenotype of the attached cells by staining with precursor and mature endothelial cell markers such as CD45, CD133, CD31 andvWF. If successful, this study could open the door to autologous endothelialistion of bioprosthetic valves in the future, reducing the need for re-operation and consequently reducing the associated health costs.

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