Table of Contents
Journal of Histology
Volume 2016, Article ID 7680701, 8 pages
http://dx.doi.org/10.1155/2016/7680701
Research Article

Histochemical and Molecular Characterization of Spongiosal Cells in Native Tissue, Two- and Three-Dimensional Cultures of Rat Aortic Valve

1Council of Scientific and Industrial Research-Centre for Cellular and Molecular Biology, Hyderabad 500007, India
2Birla Institute of Technology and Sciences, K. K. Birla Campus, Zuarinagar, Goa 403726, India

Received 7 September 2015; Revised 2 December 2015; Accepted 15 December 2015

Academic Editor: Victor H. Casco

Copyright © 2016 Vibudha Nanduri et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

The histocytochemical and molecular analysis of cells that constitute the aortic valve (AV) of the rat heart was done in this study. We have focussed on the identity of cells in the spongiosal layer of the valve by immunofluorescence studies using lineage specific markers and cytochemical staining. We have established two-dimensional (2D) cultures of cells from isolated rat AV leaflets and maintained endothelial and interstitial valvular cells (IVC) over a period of six to eight weeks. Using “passage 0” cells from 2D valvular cultures, we could reconstruct the three-dimensional (3D) valvular tissue in collagen gels that showed very similar cellular organization and marker expression profile, as that of the native tissue. Lineage specific markers in the native tissue and cell cultures were studied by Reverse Transcriptase-PCR and immunofluorescence for VCAM-I, α-SMA, collagen I, CD71, collagen II, and E-cadherin markers. This is the first report on the identification of cell lineages in the spongiosal layer of AV and the successful reconstruction of 3D valvular tissue from primary cell cultures of AV.