Reconstructed Human Cornea Produced in vitro by Tissue Engineering

Pathobiology - Tập 67 Số 3 - Trang 140-147 - 1999
Lucie Germain1, François A. Auger1, Eric Grandbois1, Rina Guignard1, Marcelle Giasson2, Hélène Boisjoly3, Sylvain L. Guérin2
1Laboratoire d'Organogénèse Expérimentale (LOEX), Hôpital du Saint-Sacrement, Québec et Département de Chirurgie, Université Laval, Sainte-Foy,
2Laboratoire d'Endocrinologie Moléculaire, Centre de Recherche du CHUL, Québec, et
3Département d'Ophtalmologie, Hôpital Maisonneuve-Rosemont, Université de Montréal, Montréal, Canada

Tóm tắt

The aim of the present study was to produce a reconstructed human cornea in vitro by tissue engineering and to characterize the expression of integrins and basement membrane proteins in this reconstructed cornea. Epithelial cells and fibroblasts were isolated from human corneas (limbus or centre) and cultured on plastic substrates in vitro. Reconstructed human corneas were obtained by culturing epithelial cells on collagen gels containing fibroblasts. Histological (Masson’s trichrome staining) and immunohistological (laminin, type VII collagen, fibronectin as well as β<sub>1</sub>, α<sub>3</sub>, α<sub>4</sub>, α<sub>5</sub>, and α<sub>6</sub> integrin subunits) studies were performed. Human corneal epithelial cells from the limbus yielded colonies of small fast-growing cells when cultured on plastic substrates. They could be subcultured for several passages in contrast to central corneal cells. In reconstructed cornea, the epithelium had 4–5 cell layers by the third day of culture; basal cells were cuboidal. The basement membrane components were already detected after 3 days of culture. Integrin stainings, except for the α<sub>4</sub> integrin, were also positive after 3 days. They were mostly detected at the epithelium-stroma junction. Such in vitro tissue-engineered human cornea, which shows appropriate histology and expression of basement membrane components and integrins, provides tools for further physiological, toxicological and pharmacological studies as well as being an attractive model for gene expression studies.

Từ khóa


Tài liệu tham khảo

10.1006/excr.1994.1300

10.1002/(SICI)1097-4652(199705)171:2<179::AID-JCP8>3.0.CO;2-L

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Thông tin xuất bản

Pathobiology

Tập 67 Số 3

140-147

Thông tin tác giả