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Primary hepatoma carcinoma cell culture medium, primary hepatoma carcinoma cell culture method and application

A technology of liver cancer cells and culture methods, which is applied to the culture of primary liver cancer cells, and the cells are used in the field of drug efficacy evaluation and screening, which can solve problems such as the inability to meet the requirements of rapid clinical detection and the long culture cycle of stable cell lines.

Active Publication Date: 2021-09-10
PRECEDO PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, using the conditioned medium (F Medim) recorded in this literature has a long culture period for stable cell lines screened in vitro, which cannot meet the needs of rapid clinical testing.

Method used

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  • Primary hepatoma carcinoma cell culture medium, primary hepatoma carcinoma cell culture method and application
  • Primary hepatoma carcinoma cell culture medium, primary hepatoma carcinoma cell culture method and application
  • Primary hepatoma carcinoma cell culture medium, primary hepatoma carcinoma cell culture method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1 Acquisition of primary liver cancer cells and treatment of trophoblast cells

[0040] 1. Collection of Liver Cancer Tissue Samples

[0041] Liver cancer tissue samples were collected within half an hour of surgical resection or puncture in explained and consented liver cancer patients. In a sterile environment, cut the liver cancer tissue sample lcm from the non-necrotic part 3 Above, place it in 15mL DMEM / F12 medium pre-cooled in a refrigerator at 4°C, and mark it.

[0042] 2. Processing of Liver Cancer Tissue Samples

[0043] (1) Take the tissue in a biological safety cabinet and place it in a 100mm culture dish (Corning, 430167), remove the tissue with blood, and use DMEM / F12 medium (manufacturer: Corning) supplemented with 1% streptomycin- Penicillin solution (hereinafter referred to as double antibody solution, purchased from Corning Company of the United States, wherein, the concentration of streptomycin in the mother solution is 10mg / ml, and the conc...

Embodiment 2

[0058] Example 2 Component optimization of primary liver cancer cell culture medium

[0059] 1% double antibody solution (purchased from Corning Company, wherein the concentration of streptomycin in the mother solution is 10mg / ml, and the concentration of penicillin is 10000U / ml) added to DMEM / F12 medium, 10% fetal bovine serum (purchased from Gibco Company) and 0.1% Primocin solution (purchased from Invivogen Company, wherein the concentration of Primocin in the mother liquor is 50 mg / ml) was used as the basal medium.

[0060] Cytokines were added to the basal medium at concentrations according to the following Table 1 to obtain the single-factor supplemented medium of Example 2.

[0061] According to the method described in Example 1, primary liver cancer cells were obtained from surgically resected liver cancer tissue samples of 4 liver cancer patients (patient 2, patient 3, patient 7, and patient 8) who had been explained and obtained consent. Using the single factor-adde...

Embodiment 3

[0068] Example 3 Culture and identification of primary liver cancer cells

[0069] The primary hepatoma cell culture medium (HCCM) of Example 3 was prepared according to the formula in Table 2 below.

[0070] 【Table 2】

[0071] serial number Types of medium additives supplier Final concentration 1 N2 Gibico 1:100 2 epidermal growth factor R&D 10ng / ml 3 hepatocyte growth factor Sino Biological 20ng / ml 4 basic fibroblast growth factor Sino Biological 20ng / ml 5 R-spondin1 Sino Biological 250ng / ml 6 Glutamine Invitrogen 250ng / ml 7 non-essential amino acids Corning 1μM 8 insulin Sigma-Aldrich 5ng / ml 9 Y27632 MCE 10μM 10 IL-6 R&D 5ng / ml 11 DMEM / F12 medium Corning 90% (volume%) 12 fetal bovine serum Gibico 10% (volume %) 13 Hydrocortisone Sigma-Aldrich 400ng / ml 14 streptomycin-penicillin Corning 1% (volume %) 15 Primocin Inviv...

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PUM

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Abstract

The invention provides a primary hepatoma carcinoma cell culture medium and a primary hepatoma carcinoma cell culture method which are used for rapidly and stably culturing primary hepatoma carcinoma cells. The primary hepatoma carcinoma cell culture medium contains glutamine, non-essential amino acid, a basic fibroblast growth factor, a hepatocyte growth factor, IL-6, an epidermal growth factor, insulin, Y27632, an N2 additive, a B27 additive, Primocin, penicillin, streptomycin, fetal calf serum, optional hydrocortisone and optional R-spondin1. A cell model obtained by the primary hepatoma carcinoma cell culture medium and the primary hepatoma carcinoma cell culture method can be used for evaluating and screening curative effects of medicines.

Description

technical field [0001] The present invention belongs to the field of biotechnology, and specifically relates to a primary liver cancer cell culture medium and a primary liver cancer cell culture method for culturing or expanding primary liver cancer cells in vitro, and also relates to the curative effect evaluation of cultured cells in drugs and screening methods and applications. Background technique [0002] Primary culture (primary culture), also called primary culture, refers to the first culture of tumor cells or tissues directly removed from the body. The most commonly used primary cultures are tissue block culture and dispersed cell culture. Tissue block culture is to directly transplant the shredded tissue block on the wall of the culture bottle, and then culture it after adding the culture medium. Dispersed cell culture is to disperse cells by mechanical or chemical methods in tissue blocks. To isolate the most viable free cells from clinical biopsies or resected...

Claims

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Application Information

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IPC IPC(8): C12N5/09C12Q1/02
CPCC12N5/0693G01N33/5011C12N2500/32C12N2500/33C12N2501/115C12N2501/12C12N2501/2306C12N2501/11C12N2501/33C12N2501/999C12Q1/02C12N5/06
Inventor 刘青松赵明陈程王文超任涛王黎
Owner PRECEDO PHARMA CO LTD
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