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G. Anastassiou, G. Prescher, D. von der Haar, D. Meller, N. Bornfeld, G. Nareyeck; Establishment of a cell line derived from a recurrent conjunctival melanoma . Invest. Ophthalmol. Vis. Sci. 2004;45(13):1224.
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Purpose: Conjunctival melanoma is a very rare disease. Due to its rarity and the fact that most patients are not primarily treated in specialized centers the availability of material for research is very limited. Therefore, the establishment of cell lines is an important step towards basic research on that tumor. Methods: The nodular tumor was located in the bulbar conjunctiva and was the third recurrence over a period of 21 years of an epithelioid conjunctival melanoma in the same area. The tumor material was isolated immediately after surgery for cultivation. The tumor was cut into small pieces followed by trypsin treatment for several minutes. Singled tumor cells were plated out on untreated culture flasks or on culture flasks containing irradiated feeder layer cells. The Ham's F12 medium with 10% FCS was used. Immunocytochemistry for HMB45, S100 and MCSP was performed. Cell proliferation was measured with the MTT proliferationassay. The tumor karyotyp was assessed by conventional cytogenetic analysis. Results: Tumor cell counts decreased over the first two moths of cultivation and recovered thereafter beginning proliferating in a stable manner after the fifth passage (130 days). The doubling time in the last 10 passages is approximately 70 hours. The cell type is of mixed character and both spindle B and epithelioid cells are present. Immunocytochemistry was positive for HMB45 and S100 whereas MCSP was not detected. Cytogenetic analysis revealed several chromosomal aberrations. The karyotyp was hypotriploid. Conclusions: So far only one cell line originated from a primary conjunctival melanoma has been described (IPC 292, Aubert et al., Int J Cancer, 1993). This is the second cell line from a conjunctival melanoma characterized ever and the first from a recurrent conjunctival melanoma. Due to its stable and well growing pattern the cell line is suitable for experimental research.
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