Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

3D culture system of primary tumor cells, and culture method and application of 3D culture system

A culture system and tumor cell technology, applied in the 3D culture system of primary tumor cells and the field of culture, can solve problems such as harsh conditions, achieve high reliability, achieve advantageous growth, and prevent dehydration and death.

Active Publication Date: 2019-12-24
SHENZHEN YOUSHENGKANG MEDICAL LAB CO LTD
View PDF8 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Low-melting point agar culture is a good anchorage-independent growth system for screening tumor cells. It is a sign of malignant transformation of cells. Normal epithelial cells do not proliferate in agar culture, but studies have found that due to conventional low-melting point agar culture Conditions are harsh, except for a small number of tumor cell lines that can proliferate well in agar, most primary tumor cells cannot proliferate well in conventional agar culture systems

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • 3D culture system of primary tumor cells, and culture method and application of 3D culture system
  • 3D culture system of primary tumor cells, and culture method and application of 3D culture system
  • 3D culture system of primary tumor cells, and culture method and application of 3D culture system

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] This embodiment provides a 3D culture system and a preparation method of the 3D culture system, and uses the 3D culture system to cultivate primary tumor cells.

[0068] (1) Isolation and culture of tumor cells

[0069] First, primary tumor single cells are obtained from postoperatively isolated tumor tissue samples from patients:

[0070] 1. The tumor tissue isolated from patients with non-small cell lung cancer (NSCLC) after surgery was subjected to single cell isolation in a sterile environment. In order to obtain a sufficient number of viable tumor cells, take a tissue block containing more tumor tissue in the central part, and the size of the surgical sample is 5 mm × 5 mm × 5 mm; fresh tissue samples are immediately immersed in a sterile centrifuge tube filled with sample preservation solution Store at 4°C and transport refrigerated. Wherein, the formula of the sample preservation solution is: DMEM without adding serum, 10 U / mL penicillin and 100 μg / mL mitomycin...

Embodiment 2

[0096] The difference from Example 1 is that the tumor cells in the middle cell matrix of the 3D culture system are replaced by normal epithelial cells, and the normal epithelial cells are primary cells extracted from normal paracancerous tissues during pathological identification of non-small cell lung cancer. cell. The result is as image 3 and Figure 4 As shown, normal epithelial cells cannot grow normally in the 3D tumor cells.

[0097] By comparing Example 1 and Example 2, it can be seen that the 3D culture system provided by the present invention can screen out tumor cells, realize the dominant growth of tumor cells, and at the same time inhibit the growth of normal epithelial cells.

Embodiment 3

[0099] This embodiment provides a 3D culture system and a preparation method of the 3D culture system, and simultaneously uses the 3D culture system to cultivate tumor cells.

[0100] (1) The difference between the isolation and culture steps of tumor cells and Example 1 is that the composition of the digestive solution used is as shown in Table 5;

[0101] Table 5 Digestive solution formula

[0102] Reagent Final concentration Collagenase I 100U / mL Dispase II 0.6U / mL DNase I 1U / mL Hyaluronidase 100U / mL Sterile HBSS (Ca 2+ &Mg 2+ )

make up

[0103] (2) Preparation of the lower layer culture medium: first prepare 2% low-melting point agarose gel, weigh 2g of low-melting point agarose, put it into a 250mL sterile blue cap bottle, and add 100mL ultrapure water to dissolve it, after autoclaving Place in a water bath at 45°C to keep it in a melted state;

[0104] Take the 2% agarose gel preheated in a water bath at 45°C and...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
concentrationaaaaaaaaaa
concentrationaaaaaaaaaa
Login to View More

Abstract

The invention provides a 3D culture system of primary tumor cells, and an application of the 3D culture system. The 3D culture system comprises lower layer gelatin, a middle layer cell substrate and an upper layer culture medium, wherein the middle layer cell substrate comprises improved agarose gel, a condition culture medium containing addition factors and tumor cells; the improved agarose gel comprises low melting-point agarose, polylysine and an extracellular matrix; the lower layer gelatin comprises agarose gel and a conditioned medium; and the upper layer culture medium is the conditionculture medium containing the addition factors. The 3D culture system provided by the invention can screen tumor cells and normal epithelial cells, besides, can realize preferential growth of the tumor cells, is high in culture success rate, can really reflect the properties of entire tumor cell groups, and can accurately represent the growth condition of in vivo tumors, and after the 3D culture system is used for medicine susceptibility detection, the obtained results are high in reliability, and clinical promotion is facilitated.

Description

technical field [0001] The present invention relates to the field of drug sensitivity detection of tumor individualized drug function, in particular to a 3D culture method for primary tumor cells, and in particular to a 3D culture system for primary tumor cells, a culture method and application thereof. Background technique [0002] Malignant tumor is a kind of disease with very high morbidity and mortality, which seriously endangers human life and health. It is one of the main diseases leading to disability and premature death. In the age group of 35-59, malignant tumor is the leading cause of death. The first one. Current treatments for malignant tumors include surgery, radiotherapy, chemotherapy, endocrine therapy, and immunotherapy. [0003] Chemotherapy, as a systemic treatment, plays a very important role in the treatment because it can widely kill tumor cells in patients and improve the survival rate and duration of patients. Although chemotherapy plays a very impor...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09C12Q1/02
CPCC12N5/0693G01N33/5011C12N2513/00C12N2533/76C12N2533/70C12N2533/52C12N2533/54C12N2533/80C12N2533/32C12N2500/32C12N2501/115C12N2500/38C12N2500/44C12N2500/24
Inventor 喻德华肖艳归
Owner SHENZHEN YOUSHENGKANG MEDICAL LAB CO LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com