Objective To isolate and purify the melanoma stem cells (MSC) in choroidal melanoma OCM-1 cells. Methods OCM-1 cells were resuscitated, and after cultured in standard Dubecco's modifided Eagle's medium (DMEM)/F12, they were cultured in serum-free medium (SFM). The cultured MSC were isolated and purified, and the positive rate of CD133, the specific markers of neurostem cells, was observed by flow cytometry (FCM). The 6th generation of the cells were stained by musashi-1 immunocytochemistry, and the rate of the positive cells was observed under the microscope. Results After the Adherent OCM-1 cells cultured in SFM, the number of the adherent number decreased obviously. The cells at the 6th generation grew as the suspended gobbets, which represented the typical grow manner of the stem cells. Positive CD133 could be found in the cells of different generations, which was 2.5％, 21.7％, and 57.8％ in the non-isolated OCM-1 cells, the 1st generation of isolated cells, and the 2nd generation cells, respectively. The positive rate of CD133 in the cells at the sixth generation was 79.8% with b positive expression of musashi-1. Conclusion MSC is in the human choroidal melanoma OCM-1 cells. The suspended stem cells may be purified by limited differentiation and serial passage in SFM. (Chin J Ocul Fundus Dis, 2007, 23: 87-90)

Citation： LIU Bin,LI YongPing,PENG Zhan. Isolation and purification of melanoma stem cells in human choroidal melanoma OCM-1 cells. Chinese Journal of Ocular Fundus Diseases, 2007, 23(2): 87-90. doi: Copy