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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

Pending Publication Date: 2022-05-10
THE WEST CHINA SECOND UNIV HOSPITAL OF SICHUAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the invention patent with publication number CN112980768A provides a method for establishing a new type of liver fibroblast cell line, which is derived from liver tumor-related fibroblasts of patients with liver metastases from colon cancer, it only provides primary cells that have been passaged to the third generation Basic identification

Method used

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  • Mouse fibroblast tumor cell strain HXLyAF-KBM and application thereof
  • Mouse fibroblast tumor cell strain HXLyAF-KBM and application thereof
  • Mouse fibroblast tumor cell strain HXLyAF-KBM and application thereof

Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention discloses a mouse fibroblast tumor cell strain HXLyAF-KBM and application thereof.The cell strain is derived from the bone marrow microenvironment in a human lymphoma cell nude mouse transplantation tumor model, the cell name of the cell strain is mouse lymphoma related fibroblast tumor cells HXLyAF-KBM, the preservation number is CCTCC NO: C2021106, the preservation date is December 9, 2021, and the preservation number is CCTCC NO: C2021106. The preservation unit is China Center for Type Culture Collection. The cell strain is a bone marrow-derived lymphoma-related mouse fibroblast tumor cell strain established for the first time at home and abroad, and can be applied to construction of a mouse fibroblast tumor animal model, a malignant bone marrow microenvironment model, a 3D tumor culture system and organoid system model, and a bone marrow microenvironment and lymphoma progress and drug resistance relationship model.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and specifically relates to a mouse fibroblast tumor cell line HXLyAF-KBM and an application thereof. Background technique [0002] The tumor microenvironment interacts with tumor cells to affect the occurrence, development and prognosis of tumors. Cancer-associated fibroblasts (CAFs) are the main cell components that constitute the microenvironment of solid tumors. They have high heterogeneity, plasticity and multi-directional differentiation potential. It plays a complex and important role in the development, metastasis, chemotherapy resistance and malignant outcome. CAFs are a kind of highly heterogeneous cells. At present, it is believed that the precursor cells of CAFs mainly come from 7 sources: bone marrow mesenchymal stem cells, intrinsic fibroblasts in tissues, astrocytes, epithelial-mesenchymal transition, endothelial cells, etc. Cell-to-mesenchymal transition, fibroblasts and oth...

Claims

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Application Information

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IPC IPC(8): C12N5/09C12N5/077A01K67/027
CPCC12N5/0693C12N5/0656A01K67/0271A01K2227/105A01K2267/0331
Inventor 顾玲徐郡李圆圆刘瀚旻
Owner THE WEST CHINA SECOND UNIV HOSPITAL OF SICHUAN
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