The Establishment Of A Nude Mice Model Transplanted With Human MDS Cell Line SKM-1 Cells

Objective: To explore the establishment of a nude mice model transplanted with human MDS cell line SKM-1 cells.Methods: We routinely cultured MDS cell line SKM-1 cells in vitro, and made for mono-cell suspension (with 1×108/ml cell density and in exponential growth behavior) and subcutaneously implanted on 4-week-old BALB/CA-nude mice respectively. All the nude mice were divided into 3 groups. The first group included 10 mice, and they were subcutaneously implanted with 0.1ml mono-cell suspension on their right scapulas, it was about 1×107 cells. The second group included 15 mice, and they were subcutaneously implanted with 0.2ml mono-cell suspension on their right scapulas, it was about 2.0×107 cells. These mice were totally-body irradiated with 400 cGy (5-MeV photons generated by a linear accelerator at a dose rate of 200 cGy/min) before being implanted within 24h. We dissected the mice with the subcutaneous tumor in sterile environment. Then we took out the tumor and made the tumor homogenate. We routinely cultured the tumor cells in vitro, and made for mono-cell suspension (with 1×108/ml cell density and in exponential growth behavior) and subcutaneously implanted on the third group of mice respectively. The third group also included 15 mice, and they were subcutaneously implanted with 0.2ml mono-cell suspension on their right scapulas, it was about 2.0×107 cells. These mice were totally-body irradiated with 400cGy before being implanted within 24h. The results showed that the rate of the subcutaneous tumor formation of the first group was 0 (0/10), and the rate of the subcutaneous tumor formation of the second group was 13.3%(2/15), and the rate of the subcutaneous tumor formation of the third group was 53.3%(8/15). The rate of the subcutaneous tumor formation of the group 3 was obviously higher than the rate of group 1 (P=0.005) and the rate of group 2(P=0.020).Subsequently, the biological characteristics of the subcutaneous tumor cells were evaluated by the methods of cell morphology, histopathology, immunology by flow cytometer, Chromosome analysis and immunohistochemistry in nude mice model.Results: The result showed that the necrotic tissue was seen in the center after HE stain at high magnification microscopy, and the tumor cells were surrounded by inflammatory cells. Tumor cells grew irregularly, cell nucleuses were deeply stained, and the ratio of nucleus and plasma was high. The CD13: 38.41%, HAL-DR: 45.06% were similar to the cell line SKM-1 and CD3, CD5, CD7, CD14 lower by immunologic analysis by flow cytometry. The chromosome del(1q), del(1p), del(2p), del(4p), del(5q), del(6q), der(9)add(q34)del(p21), add(15p) and +1, +2, +3, +4, +5, +7, -8, +9, +10, +11, +13, -14, +15, +16, +17, +18, +21, +22 were observed. The over-expression of P53 protein was detected in tumor cells of MDS nude mice model. Our results showed that the biological characteristics of the tumor cells of MDS nude mice model were consistent with human MDS cell line SKM-1.Summary: A nude mice model transplanted with human MDS cell line SKM-1 cells was successfully established, which would be used for studying the pathogenesis of MDS and exploring the mechanism of drug treating MDS in vivo.