疾病动物模型,急性早幼粒白血病 z-bio 2021-03-08 248

　　"Severe combined immunodeficiency mice are CB-17 inbred mice and have been identified in Bosma, USA. The 16th autosomal recessive mutation (SCID) of the mouse is a mutant mouse 1. Homozygous SCID small The mouse gene almost completely loses the function of mouse T and B lymphocytes, and has been widely used in immunology and oncology studies of such mice. Acute panmyelocytic leukemia (APL) may pass the differentiation agent all-trans It is often a granular system that differentiates positively due to the effect of retinoic acid. The cause of APL is still unclear. We will study the differentiation mechanism of APL and it is very important to establish a stable animal model.

　　1 Materials and methods

　　1.1 Information

　　1.1.1 Human leukemia cell line Human acute premyeloid cell line HL-60 cell line introduced from the University Cancer Institute in 10% bovine serum (Hangzhou Sijiqing) RPMI-1640 medium, Central and South China. (Gibco product), 5% CO2, 37°C culture.

　　1.1.2 "SCID "mice" are 16 males (purchased from Shanghai Experimental Animal Center, Chinese Academy of Sciences), 3-4 weeks old. The mice are raised in a laminar flow rack with a covered mouse box (in line with SPF standards) The efficiency of filtering indoor air is moderate, while the drinking water is disinfected redistilled water with a small amount of salt. Vitamin C, eggs and debris are added to the standard pellet feed, and all items in contact with the rat are destroyed Bacteria. Divide them into two groups, each with 8 animals, and each animal was directly inoculated with 5×106 cells intraperitoneally or intravenously. 1.2 Method 1.2.1 1.2.1 SCID mice after transplantation were not treated before transplantation HL-60 cells in the logarithmic growth phase are used for transplantation. Each mouse is injected with 5×106 cells in the tail vein or intraperitoneally. 1.2.2 Peripheral blood white blood cell count and classification Before vaccination and after vaccination 1 , 2, 3, and 4 weeks to collect and test tail vein blood.

　　1.2.3 Morphological observation Collect peripheral blood, stain with Giemsa, observe cell morphology under oil microscope, and count 200 per piece of rod-shaped nucleus and lobular nucleus according to early immature, mid- juvenile, late immature. cell. Calculate the classification count, the percentage of promyelocytic cells.

　　1.2.4 Carry out pathological examination of dying animals, carry out pathological examination, collect liver, spleen, kidney, bone, heart, lung, muscle, testis and tumor body, fix with 10% formaldehyde for more than 48 hours, paraffin section, and examine under microscope.

　　1.2.5 Chromosome karyotype analysis Use standard chromosome staining techniques to culture HL-60 cells in vitro, and sample abdominal tumor tissue cells and SCID mouse blood cells according to conventional methods. Culture the RPMI-1640 medium containing abdominal tumor tissue cells, SCID mouse blood cells and 10% fetal bovine serum in a CO2 incubator at 37°C and 5% CO2 saturated humidity for 45-66 hours, and the final concentration is 0.4 cor . It's a block. Metaphase cells are then collected and subjected to hypotonicity, fixation, instillation, G-banding and microscopic examination.