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Z.M. Correa, S. Bakalian, J. Cools–Lartigue, S. Cruess, V. Saraiva, M.N. Burnier, Jr; The Role of Fine Needle Aspiration Biopsy in Cell Culture of Uveal Melanoma . Invest. Ophthalmol. Vis. Sci. 2005;46(13):3359.
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Purpose: To assess the reliability of fine–needle aspiration biopsy (FNAB) in harvesting a sufficient number of viable melanoma cells from an animal model of uveal melanoma, with the goal of establishing a cell culture and maintaining a melanoma cell line. Methods: Twelve New Zealand albino rabbits had their right eye surgically inoculated with uveal melanoma cell line 92.1. The animals were immunosupressed with cyclosporine–A using a dose schedule previously published by The Henry C. Witelson Ocular Pathology Laboratory, McGill University, Montreal, Canada. The animals were followed for 12 weeks. Intraocular tumor growth was monitored weekly by indirect ophthalmoscopy. After the fourth week, one rabbit per week was biopsied prior to sacrifice. The biopsy procedure consisted of a fine–needle aspiration using a sharp 25–gauge, 1–inch long needle, monitored under indirect ophthalmoscopy. In all cases, 2 separate aspirates were made from different areas of the tumor and flushed into individual 15 ml tubes containing cell culture medium. The cells were stored in liquid nitrogen after two weeks or when there was a minimum of 1 million cells. Cells were defrosted for immuno–staining with HMB–45 in order to confirm their melanoma origin. Results: Cell growth was observed in the samples harvested from 11 out of the 12 animals inoculated with uveal melanoma. At the time of initial cell culture, each fine needle biopsy resulted in the establishment of several foci of cell growth. Cellular morphology of the cultured fine needle aspirates closely resembled that of the 92.1 cell line originally inoculated into the eyes of the rabbits. All cells were viable after thawing from storage in liquid nitrogen and immunoassayed positively for HMB–45. Conclusions: Fine needle aspiration biopsy appears to be a reliable method of harvesting melanoma cells from primary intraocular tumors of an immunosuppressed animal model. It was shown that these cells could be re–cultured for use in potential in vitro assays. Proving the ability to establish a cell culture from FNAB harvested uveal melanoma cells of an animal model opens many future possibilities including a similar use in patients with this tumor prior to undergoing conservative treatment.
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