Collect logarithmic tumor cells, use serum-free matrix 10.0ml, centrifuge at 1500 rpm for 3min, wash 3 times in a row, and finally mix with serum-free matrix, calculate the number of tumor cells using a hemocytometer (average 5 out of 5) The cell count in the square × 104 is the number of tumors/ml). After calculating the total number of tumor cells, the density of tumor cells was adjusted to 4×106 cells/ml. Each mouse was inoculated under the arm with 50 ul (ie 2×105 tumor cells), and the tumor nodules could be palpated in about 2 weeks. Generally, mice of 6-8 weeks are selected. The number of different tumor cells inoculated is different from that of animals. For example, LL/2, B 16, Hep can be inoculated into C57 and BAL B/C mice, NS-1, EL-4, C26, Meth A can inoculate BAL B/C mice, H22 inoculate Kunming mice. After the tumor is inoculated, the small nodules can be palpated. The vernier caliper (unit: mm) is measured every 3 days by a vernier caliper for at least one month.
Pay attention to the design of different experimental groups and control groups. The number of animals in each group is generally 5-10. After 6-8 weeks of tumor inoculation, the tumor of the mouse can grow to a diameter of 15-20mm (that is, the mouse will be on the verge of death. ), blood can be taken from the eyeball, serum can be separated and preserved, mice can be sacrificed, tumor tissue can be taken for photography, part of the tumor tissue can be taken for frozen tissue section (or stored at -80℃), corresponding immunohistochemical staining can be performed, and part of the tumor can be taken. The tissue was fixed with 3% neutral formaldehyde, embedded in paraffin, and stained with conventional HE.