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CM Akor, R Campbell, H Yang, HE Grossniklaus; Effect of Angiostatin on the in vitro Melanoma Invasion . Invest. Ophthalmol. Vis. Sci. 2002;43(13):1121.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: To characterize invasive activity in vitro of murine melanoma cell lines and the effect of angiostatin on invasiveness. Methods: Matrigel© basement membrane matrix invasion chambers were filled with 500µL bicarbonate-based culture media. The chambers were rehydrated for 2h at 37°C and the media was removed. B16LS9 and Queens melanoma cells were washed with PBS, trypsinized, centrifuged and transferred to DMEM in concentrations of 1X105 in 100µL. Each 100µL solution was added to the Matrigel© invasion chamber well and 450µL of DMEM was placed in the bottom of each chamber. Each melanoma cell line was added to a well alone, with 50nM glu-plasminogen, with 250nM angiostatin or with 50nM glu-plasminogen plus 250nM angiostatin. Each concentration was repeated in triplicate. The cells were incubated at 37°C for six hours and then non-invading cells were removed with a cotton swab. The invading cells were stained with Diff-Quick. Five high-power fields were counted using a micrometer. Results: The results showing number of invading cells are shown below. Results View OriginalDownload SlideView OriginalDownload Slide Conclusions: B16LS9 and Queens melanoma cells invade Matrigel© synthetic basement membranes. Plasminogen enhances invasion of B16LS9 cells, and angiostatin reduces invasion of B16LS9 and Queens cells.
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