HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Add to Personal Folders Download to citation manager Author home page(s): Helmut Gulbins Bruno M. Meiser Bruno Reichart Sabine Daebritz Permission Requests Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Gulbins, H. Articles by Daebritz, S. Search for Related Content PubMed PubMed Citation Articles by Gulbins, H. Articles by Daebritz, S. Related Collections Related Article

Ann Thorac Surg 2005;79:2119-2126
© 2005 The Society of Thoracic Surgeons

---

New technology

Seeding of Human Endothelial Cells on Valve Containing Aortic Mini-Roots: Development of a Seeding Device and Procedure

Department of Cardiac Surgery, University Hospital Grosshadern, Munich, Germany

Accepted for publication May 6, 2004.

^_* Address reprint requests to Dr Gulbins, Department of Cardiac Surgery, University Hospital Ulm, Steinhövelstr 9, D-89070 Ulm, Germany (E-mail: helmut.gulbins{at}medizin.uni-ulm.de).

PURPOSE: Complete covering of an artificial valvular scaffold with endothelial cells may prevent thromboembolic complications and lead to an excellent biocompatibility. For this purpose, we developed a seeding device for reproducible cell seeding on valve containing aortic roots.

DESCRIPTION: Human endothelial cells and fibroblasts were obtained from saphenous vein pieces. Cryopreserved aortic roots (n = 25) were put into an especially developed tube, set on a rotator, and incubated with the cell suspension. The device rotated in two axes (sagittal and axial), ensuring slight movements of the leaflets. The rotation alternated with resting periods, allowing cell attachment to the surface. Different resting periods were tested (groups 1, 2, and 3 were 30, 45, and 60 min, respectively; n = 5 each). Total incubation time was 24 hours followed by further culturing for 6 days. In two further groups (groups 4 and 5; n = 5 each), a modified inlay was used to allow the cell suspension to flow around the entire graft. In group 4 the grafts were again incubated with human endothelial cells; however, in group 5 pre-seeding with autologous fibroblasts was done in addition. Immunohistochemical staining with antibodies against factor VIII, CD31, laminin, collagen IV, and CD90 were done, and scanning electron microscopy was done after initial seeding and after 6 days in culture.

EVALUATION: Seeding resulted in homogenous cell layers on the luminal surface of the free walls in all groups. With resting periods of 45 minutes, these results were also obtained on the leaflets, whereas the other resting times resulted in defects of the endothelial cell layer on the cusps. After 6 days under culture conditions, the endothelial cell layers were confluent and viable, with the exception of the leaflets in group 1. With the modified inlay (groups 4 and 5), confluent cell layers were also achieved on the outer surface. In group 5 pre-seeding with autologous fibroblasts resulted in enhanced synthesis of extracellular matrix proteins, as was demonstrated with immunohistochemical staining for collagen IV and laminin.

CONCLUSIONS: With this newly developed seeding device, confluent cell layers on valve containing aortic roots were reproducibly achieved. The technique enables further experimental research and even clinical application.

Related Article

Cardiovascular Tissue Engineering Therapy: So Near, So Far?

Gilbert H.L. Tang, Shafie Fazel, Richard D. Weisel, Glen S. Van Arsdell, and Ren-Ke Li
Ann. Thorac. Surg. 2005 79: 1831-1833. [Extract] [Full Text] [PDF]

This article has been cited by other articles:

				P. M. Dohmen and W. Konertz Seeding human endothelial cells on complex three-dimensional scaffolds. Ann. Thorac. Surg., May 1, 2006; 81(5): 1942 - 1942. [Full Text] [PDF]

				Reply. Ann. Thorac. Surg., May 1, 2006; 81(5): 1942 - 1943. [Full Text] [PDF]

				G. H.L. Tang, S. Fazel, R. D. Weisel, G. S. Van Arsdell, and R.-K. Li Cardiovascular Tissue Engineering Therapy: So Near, So Far? Ann. Thorac. Surg., June 1, 2005; 79(6): 1831 - 1833. [Full Text] [PDF]