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Additive for primary culture of cancer cells as well as culture medium and application thereof

A primary culture and culture medium technology, applied in the direction of tumor/cancer cells, animal cells, vertebrate cells, etc., can solve the problems of ineffective inhibition and removal, cumbersome establishment of culture system, inconvenient operation, etc., and achieve easy quality Control the system, reduce the difficulty of large-scale preparation, and realize the effect of commercialization

Pending Publication Date: 2022-08-05
南宁云幂方生物医药技术有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the process of culturing primary tumor cells in the existing serum-free medium, it is still necessary to introduce irradiated trophoblast cells from xenogeneic sources into the culture system, which makes the establishment of the culture system too cumbersome and inconvenient to operate. It also poses challenges to the standardization and large-scale production of the kit, and it is difficult to achieve mass production. At the same time, it cannot effectively inhibit and remove fibroblasts, the main confounding component in primary tumor cells.

Method used

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  • Additive for primary culture of cancer cells as well as culture medium and application thereof
  • Additive for primary culture of cancer cells as well as culture medium and application thereof

Examples

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Effect test

Embodiment 1

[0047] This example provides a medium for primary culture of cancer cells, including 200 mL of complete DMEM medium and 100 mL of F12 nutrient mixture; the F12 nutrient mixture includes 2 mg of hydrocortisone, 1 mg of EGF, 2 mg of insulin, 1 mg of amphotericus Supplement B, 3 mg of gentamicin and 5 mg of supplements for primary culture of cancer cells;

[0048] The additives for primary culture of cancer cells include cholera toxin, R-spondin, Y-27632, and the mass ratio is 1:2:2.

Embodiment 2

[0050] This example provides a medium for primary culture of cancer cells, including 400 mL of complete DMEM medium and 100 mL of F12 nutrient mixture; the F12 nutrient mixture includes 4 mg of hydrocortisone, 3 mg of EGF, 5 mg of insulin, 5 mg of amphoteric Supplement B, 5 mg of gentamicin and 5 mg of supplements for primary culture of cancer cells;

[0051] The additives for primary culture of cancer cells include cholera toxin, R-spondin, Y-27632, and the mass ratio is 1:3:5.

Embodiment 3

[0053] This example provides a medium for primary culture of cancer cells, including 300 mL of complete DMEM medium and 100 mL of F12 nutrient mixture; the F12 nutrient mixture includes 3 mg hydrocortisone, 2 mg EGF, 3.5 mg insulin, 3.5 mg Amphotericin B, 4 mg of gentamicin and 5 mg of supplements for primary culture of cancer cells;

[0054] The additives for primary culture of cancer cells include cholera toxin, R-spondin, Y-27632, and the mass ratio is 1:2.5:3.5.

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Abstract

The invention provides an additive for primary culture of cancer cells as well as a culture medium and application thereof, and belongs to the technical field of medical treatment. The additive for the primary culture of the cancer cells is characterized by comprising at least three of cholera toxin, R-spondin, A-83-01 and Y-27632. The culture medium for primary culture of cancer cells comprises a complete DMEM culture medium and an F12 nutrient mixture, the F12 nutrient mixture comprises at least three of cholera toxin, R-spondin, A-83-01 and Y-27632, and the F12 nutrient mixture comprises at least three of cholera toxin, R-spondin, A-83-01 and Y-27632. By optimizing and screening a culture method and a culture medium formula, a culture system independent of trophoblast cells is developed, and the success rate is high, so that the large-scale preparation difficulty of the culture medium is greatly reduced, meanwhile, a stable quality control system is easier to establish, and commercialization is realized.

Description

technical field [0001] The present invention relates to the technical field of detection kits, in particular to an additive for primary culture of cancer cells, a culture medium and use thereof. Background technique [0002] Because the primary tumor cells have just been separated from the tissue, the biological characteristics have not changed greatly, and the biological characteristics of the tumor in the original state can be well maintained. Due to differences in genetic backgrounds, the pathogenesis of different patients and the sensitivity to treatment have individual differences. Therefore, the primary culture of autologous tumor cells is an important means to study the pathogenesis of tumors and the molecular biological characteristics of tumor cells. [0003] Artificially simulating the living environment of tumor cells in vitro is the basis of primary tumor cell culture. In addition to environmental factors such as sterility, temperature, pH value, and humidity, th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09C12N5/071C12Q1/02
CPCC12N5/0693C12N5/0682G01N33/5011C12N2501/01C12N2501/415C12N2501/15C12N2501/727C12N2501/39C12N2501/11C12N2501/33C12N2503/02G01N2500/10C12N2509/00C12N2509/10
Inventor 武瑞琴王正远杨海
Owner 南宁云幂方生物医药技术有限责任公司