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Application of serum-free culture medium in culturing cancer tissue origin spheroids

A serum-free medium and culture medium technology, applied in the field of cell culture, can solve the problems of limited proliferation, large cell loss and slow proliferation of CTOS culture, and achieve stable and controllable culture conditions

Pending Publication Date: 2021-04-06
GUANGZHOU DARUI BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, there are still some deficiencies in the existing methods, such as the large amount of cell loss during the preparation of CTOS; the selection of CTOS mainly relies on manual operation, which is time-consuming, laborious, and low-throughput; the proliferation of CTOS culture is limited and the proliferation is slow. Limited samples are collected, and its clinical application still has great limitations

Method used

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  • Application of serum-free culture medium in culturing cancer tissue origin spheroids
  • Application of serum-free culture medium in culturing cancer tissue origin spheroids
  • Application of serum-free culture medium in culturing cancer tissue origin spheroids

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] Embodiment 1 A kind of serum-free culture medium

[0074] Each component and its final concentration: basal medium DMEM / F12; BSA, 1.55% (w / v); Y27632, 10μM; B27 (50×), 2% (v / v); Heregulin, 10ng / ml; EGF , 50ng / ml; bFGF, 8ng / ml; Activin A, 10ng / ml; Nicotinamide, 10mM; NEAA (100×), 1% (v / v); Trace Elements A (1000×), 0.1% (v / v); TraceElements B (1000×), 0.1% (v / v); Trace Elements C (1000×), 0.1% (v / v); 2-mercaptoethanol, 0.1 mM; Primocin, 100 μg / ml.

Embodiment 2

[0075] Embodiment 2 A kind of serum-free culture medium

[0076] Components and their final concentrations: basal medium DMEM / F12; BSA, 0.1% (w / v); Y27632, 1 μM; B27 (50×), 0.1% (v / v); Heregulin, 1ng / ml; EGF , 1ng / ml; bFGF, 1ng / ml; Activin A, 1ng / ml; Nicotinamide, 1mM; NEAA (100×), 0.1% (v / v); Trace Elements A (1000×), 0.01% (v / v); TraceElements B (1000×), 0.01% (v / v); Trace Elements C (1000×), 0.01% (v / v); 2-mercaptoethanol, 0.01 mM; Primocin, 50 μg / ml.

Embodiment 3

[0077] Embodiment 3 A kind of serum-free culture medium

[0078]Each component and its final concentration: basal medium DMEM / F12; BSA, 10% (w / v); Y27632, 50μM; B27 (50×), 10% (v / v); Heregulin, 100ng / ml; EGF , 200ng / ml; bFGF, 100ng / ml; Activin A, 100ng / ml; Nicotinamide, 100mM; NEAA (100×), 10% (v / v); Trace Elements A (1000×), 1% (v / v); TraceElements B (1000×), 1% (v / v); Trace Elements C (1000×), 1% (v / v); 2-Mercaptoethanol, 1 mM; Primocin, 500 μg / ml.

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Abstract

The invention discloses a serum-free culture medium, which comprises a basal culture medium, serum albumin, an ROCK inhibitor, B27, Herbulin, Activin A, EGF, bFGF, nicotinamide, NEAA, Trace Elements A, Trace Elements B, Trace Elements C and 2-mercaptoethanol. A convenient, rapid and stable culture method of the cancer tissue origin spheroid is established by utilizing the serum-free culture medium, the culture medium composition and the preparation method in the original culture method are optimized, and a stable and controllable serum-free culture scheme is provided; the problems that in an existing three-dimensional culture scheme, the primary tumor cell sample size is insufficient, and the proliferation effect is limited are solved, and model support is provided for preparing a CTOS biological sample library and conducting personalized medicine evaluation and screening on cancer patients.

Description

technical field [0001] The invention relates to the technical field of cell culture, and more specifically relates to the application of a serum-free medium in culturing spheroids of cancer tissue origin. Background technique [0002] Malignant tumor is a common disease that threatens human health. It ranks first among the three major causes of death in human diseases, accounting for a quarter of all disease deaths. With the development of the economy and the improvement of living standards, the incidence of tumors is increasing year by year. It is estimated that by 2030, the number of morbidity and death from tumors in the world will reach 26.4 million and 17 million, respectively. The treatment of tumors mainly relies on technical means such as surgery, radiotherapy and chemotherapy, targeted therapy and immunotherapy. Against the backdrop of the global development of precision medical technology, the development of new anti-cancer drugs is becoming increasingly important...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09
CPCC12N5/0693C12N5/0679C12N5/0631C12N2501/727C12N2501/115C12N2501/11C12N2500/44C12N2500/38C12N2501/998C12N2513/00C12N2500/90C12N2533/54
Inventor 万勇徐中华何志华李明
Owner GUANGZHOU DARUI BIOTECH