Mouse fibroblast tumor cell strain HXLyAF-KBM and application thereof
A fibroblast and tumor cell line technology, applied to the mouse fibroblast tumor cell line HXLyAF-KBM and its application field
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Embodiment 1
[0025] Example 1: Isolation and establishment of mouse fibroblast tumor cell line HXLyAF-KBM
[0026] (1) The human anaplastic large cell lymphoma cell Karpas299 in the logarithmic growth phase was divided into 8×10 6 Each / 100µl / mouse, inoculated subcutaneously in the nude area of female Balb / c (nu / nu) mice (8 weeks old, body weight 25.41g), the weight of the mice decreased progressively 67 days after inoculation, and the mice were harvested when the body weight was 20.02g The mice were sacrificed and dissected, and the bone marrow in the bilateral femur and tibia was taken to make a cell suspension, and the cells were resuspended in RPMI 1640 medium (Gibco Company) containing 10% fetal bovine serum (Thermo Company), placed at 37°C, saturated Humidity, 5% CO 2 Incubator cultivation. After 4 days, the growth of adherent cells can be seen. Remove the supernatant and suspension cells, and change the medium every 2-3 days. After 16 days, the suspension cells in the culture dis...
Embodiment 2
[0032] Example 2: Identification of growth and biological characteristics of mouse fibroblast tumor cell line HXLyAF-KBM
[0033] (1) Observation of cell morphology:
[0034] Take the HXLyAF-KBM cells in the logarithmic growth phase and observe the living cell morphology under an inverted microscope, as shown in figure 1 As shown, the cells grow adherently, and the morphology is mainly polygonal star-shaped, fusiform, large nuclei, clear nucleoli, no contact inhibition, cross-overlapped growth, and clusters when the cells are dense; pre-fixed with 3% glutaraldehyde , re-fixed with 1% osmium tetroxide, and stained with uranyl acetate and lead citrate to observe the ultrastructure of the cells under the perspective electron microscope of JEM-1400FLASH, such as figure 2 As shown, the nuclei are irregular in shape with notches, clear nuclear membrane, obvious nucleoli, concave nuclei, increased rough endoplasmic reticulum in the cytoplasm, abundant free ribosomes, endoplasmic reti...
Embodiment 3
[0044] Example 3: Construction of Mouse Fibroblast Tumor Animal Model
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