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Denise Loya, Alba Guerrero-Martinez, Eduardo Camacho-Martinez, Jorge Luis Domene Hickman, Paloma Lopez, Julio C Hernandez-Camarena, Judith Zavala, Jorge E Valdez; Molecular characterization of pterygium fibroblasts in an enhanced serial explant culture technique. Invest. Ophthalmol. Vis. Sci. 2018;59(9):142.
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To analyze and validate the stability of the pterygium fibroblastic cells obtained through a serial explant technique by immunocytochemistry analysis with two specific (ALDH3A1, PRDX2) and non-specific (pan-keratin) fibroblastic pterygium biomarkers
Pterygium tissue was obtained and cultured with a serial explant technique using extracellular matrix coating. After the primary culture, four explant transfers were done: cell morphology preservation, replication rate and growth patterns were achieved. Molecular characterization through immunocytochemistry (Marchenko and Flanagan modified method) was performed: cell fixation (4% paraformaldehyde) to preserve cell morphology, posteriorly washed and permeabilized with 0.3% Triton X-100, bovine serum albumin (BSA) was used as a blocking agent. Later, cells were stained for ALDH3A1, PRDX2 and Pan-keratin. The same procedure was implemented on the standard fibroblast cell line (NIH3T3) (control group). Photographic images were taken with a fluorescence microscope
Molecular expression of ALDH3A1, PRDX2 and pan-keratin was present and holded on positive in the primary culture as well as in the four explant transfer cell population
The use of serial explants of pterygium fibroblastic cells proved to be an achievable method to increase the number of cells obtained from a single tissue sample. Molecular stability was demonstrated throughout all the explant cell populations, suggesting that this technique is a feasible option for enhancing human pterygia fibroblasts. The use of these cells in future studies can be helpful in the evaluation of new therapeutic agents
This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.
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