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Aaron Yeung, Andrew Hopkinson, Harminder Dua; Thermolysin treated AM preserves the BM providing superior expansion and preservation of cornea epithelial cells. Invest. Ophthalmol. Vis. Sci. 2013;54(15):990.
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© ARVO (1962-2015); The Authors (2016-present)
: Amniotic Membrane (AM) has grown in popularity over the years as a substrate for epithelial expansion and therefore a potential treatment adjuvant in limbal stem cell deficiency. The AM basement membrane (BM) provides the necessary structure for cell attachment, and the AM itself is postulated to possess anti-inflammatory and anti- scarring properties. Tenascin C (TNC) is a multimodular glycoprotein that is part of the ECM located at the BM. It has the potential to engage within the microenvironment of the stem cell niches. TNC may have a role in maintaining corneal epithelial stemness. Thermolysin treated AM is a standardized method for denuding AM whilst preserving the BM. The aims of our study were to investigate whether thermolysin treated AM could provide a suitable microenvironment for expansion and preservation of epithelial cells.
Primary cells from corneo-scleral explants were cultured on intact AM and thermolysin treated AM. Relative quantification for various “stem cell associated” markers was performed using both real time PCR and immunofluorescence.
Thermolysin treated AM demonstrated greater stratification on primary cells with higher levels of TNC expression and maintaining cells with greater p63 and ABCG2 expression and lower DSG levels.
The AM could provide a suitable microenvironment similar to a stem cell niche for expansion and preservation of epithelial cells. Thermolysin treated AM preserves the BM and therefore permits superior cell to BM adhesion. Preservation of the AM BM may be the key to underlying cell interactions that maintain stem cell “stemness”.
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