Human anterior lens capsule serving as a substrate for human trabecular meshwork cells cultivation
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ORIGINAL PAPER
Human anterior lens capsule serving as a substrate for human trabecular meshwork cells cultivation Nikolaos Kopsachilis • Konstantinos T. Tsaousis Ioannis T. Tsinopoulos • Friedrich E. Kruse • Ulrich Welge-Lu¨ssen
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Received: 10 January 2012 / Accepted: 18 July 2012 / Published online: 27 July 2012 Ó Springer Science+Business Media B.V. 2012
Abstract To investigate if human anterior lens capsule is a suitable substrate for the culture of primary human trabecular meshwork (HTM) cells. Trabecular meshwork cells derived from four human donors were seeded on anterior lens capsules that were prepared from the lenses of donor eyes. Cell morphology and viability were examined at 1, 3, 5 and 7 days. Cell viability was measured based on a two-colour fluorescence assay (membrane-impermeable propidium iodide and membrane permeable Hoechst 33342). Immunocytochemistry studied Zonula occludens-1 (ZO-1), vimentin, tissue transglutaminase (tTgase) and Na?/K?-adenosine triphosphatase (Na?/K?ATPase). Morphology of the cultivated cells followed a typical model while their viability was[95 % in all cases. ZO-1 was found at the cell boundaries of the HTM-AC complex. Vimentin was located at the lateral membranes of the HTM cells. Na?/K?-ATPase was found at the basolateral membrane of the HTM cells. tTgase was also identified. Anterior lens capsule can be considered as a suitable alternative substrate for
N. Kopsachilis K. T. Tsaousis F. E. Kruse U. Welge-Lu¨ssen Department of Ophthalmology, Friedrich-Alexander University Erlangen-Nuremberg, Erlangen, Germany K. T. Tsaousis (&) I. T. Tsinopoulos 2nd Department of Ophthalmology, Medical School, Aristotle University of Thessaloniki, ‘‘Papageorgiou’’ General Hospital, 56429 Thessaloniki, Greece e-mail: [email protected]
cultivation of HTM cells and assist the expansion of existing knowledge about glaucoma pathophysiology and therapy. Keywords Trabecular meshwork Cell culture Glaucoma Anterior lens capsule
Introduction Cultivation of human cells of any kind is without doubt a challenging project. Through the development of a whole new era, described briefly by the term ‘‘Biomedical Engineering’’, cultivated cells could plausibly have practical applications, such as assisting the building process of new healthy structures able to substitute damaged ones in the human body. In addition, cultivating various types of cells can lead to the creation of an arsenal for numerous in vitro studies. In ophthalmology, there have been many attempts to culture different kind of cells as corneal epithelium and endothelium cells, retinal pigmented epithelium cells, astrocytes, keratinocytes and cells of the trabecular meshwork (Castro-Mun˜ozledo 2008; Orwin and Hubel 2000; Kobayashi et al. 2005; Tezel et al. 2001; Lin et al. 2007). The potential ultimate aim of these efforts is the in vivo application of the cultivated biomaterials in order to treat diverse ocular pathological entities (Nishida 2003). Such clinical problems could be corneal
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