Effective culturing method for body fluid derived tumor cells and application of effective culturing method

A tumor cell and culture method technology, applied in the field of tumor cell culture, can solve problems such as differences in genetic backgrounds and gene expression profiles, long culture periods, and differences in drug responsiveness

Pending Publication Date: 2018-10-26
上海赫佰生物医药科技有限公司
View PDF6 Cites 7 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the tumor cell line has the advantages of short culture cycle, easy operation, and low cost. The disadvantages are: 1) The tumor cell line is cultured under 2D conditions, which is quite different from the in vivo environment; 2) The genetics of the tumor cell line Background and gene expression profiles are quite different from clinical patient tumor cells
Its disadvantages are as follows: 1) The culture cycle is long, and it usually takes several months from tumor tissue acquisition to inoculation, subculture, and experiment; 2) It needs special immunodeficiency mice and animal culture places, which are expensive; 3) Experimental animals are sensitive to drugs. Reactivity, etc. are quite different from humans
CN102439135B discloses the culture of stem cell organoids contained in normal epithelial tissues and the 3D culture of tumor cells derived from epithelial adenoma tissue blocks, but does not involve the isolation of tumor cells from body fluids, especially

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
  • Effective culturing method for body fluid derived tumor cells and application of effective culturing method
  • Effective culturing method for body fluid derived tumor cells and application of effective culturing method
  • Effective culturing method for body fluid derived tumor cells and application of effective culturing method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Embodiment 1; The cultivation of gastric cancer patient's ascites tumor cell

[0056] 1. Extraction of tumor cells from ascites of patients with gastric cancer

[0057] 1. Collect pleural effusion samples from stage IV gastric cancer patients (the samples in this invention are taken from Changhai Hospital Affiliated to Second Military Medical University), and take 20ul microscopic examination to observe the overall condition of tumor cells or cell clusters: shape, quantity, and size.

[0058] 2. After centrifugation at 1200 rpm for 5 minutes, collect the supernatant, and filter the supernatant through a membrane filter to sterilize it, aliquot it, and inactivate it in a water bath at 50-60°C for 30 minutes.

[0059] 3. Use PBS (phosphate-buffered saline) to gently resuspend the bottom pellet, mix gently and let it stand, observe the sedimentation, and suck away the supernatant as much as possible.

[0060] 4. Resuspend the pellet with PBS, mix gently, let stand or cent...

Embodiment 2

[0070] Embodiment two; The cultivation of gastric cancer patient's cerebrospinal fluid tumor cell

[0071] 1. Extraction of tumor cells from cerebrospinal fluid of patients with gastric cancer

[0072] 1. Collect cerebrospinal fluid samples from stage IV gastric cancer patients (the samples in this invention are taken from Changhai Hospital Affiliated to Second Military Medical University), and take 20ul microscopic examination to observe the overall condition of tumor cells or cell clusters: shape, quantity, and size.

[0073] 2. After centrifugation at 1200 rpm for 5 minutes, collect the supernatant, and filter the supernatant through a membrane filter to sterilize it, aliquot it, and inactivate it in a water bath at 50-60°C for 30 minutes.

[0074] 3. Use PBS (phosphate-buffered saline) to gently resuspend the bottom pellet, mix gently and let it stand, observe the sedimentation, and suck away the supernatant as much as possible.

[0075] 4. Resuspend the pellet with PBS, ...

Embodiment 3

[0085] Embodiment three; The cultivation of ascites tumor cell of intestinal cancer patient

[0086] 1. Extraction of tumor cells from ascites of patients with intestinal cancer

[0087] 1. Collect ascites samples from patients with stage IV bowel cancer (the samples in this invention are taken from the Cancer Hospital Affiliated to Fudan University), and take 20ul microscopic examination to observe the overall condition of tumor cells or cell clusters: shape, quantity, and size.

[0088] 2. After centrifugation at 1200 rpm for 5 minutes, collect the supernatant, filter the supernatant through a membrane filter and sterilize it, then aliquot it, and inactivate it in a water bath at 50-60°C for 30 minutes.

[0089] 3. Use PBS (phosphate-buffered saline) to gently resuspend the bottom pellet, mix gently and let it stand. After observing the sedimentation, suck away the supernatant as much as possible.

[0090] 4. Resuspend the pellet with PBS, mix gently, let stand or centrifug...

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

No PUM Login to view more

Abstract

The invention relates to a culturing method for body fluid derived tumor cells and a tumor medium. The method comprises the steps of enriching body fluid tumor cells; mixing the body fluid tumor cellswith an extracellular matrix; and in the presence of a basic medium, adding corresponding pleural effusion, ascites or cerebrospinal fluid and the like to carry out 3D cell culture. By the method, apatient derived tumor analog (PDTA) can be obtained effectively, and a novel detecting tool is provided for detection of sensitivity of clinical tumor patients to drugs and measurement of in-vitro metabolic stability, metabolic profiling, toxicity and the like of drug candidates.

Description

technical field [0001] The invention relates to the field of culturing tumor cells derived from body fluids, in particular to a culturing method for various pleural effusion, ascites or cerebrospinal fluid tumor cells, a tumor cell culture medium and applications thereof. Background technique [0002] Currently, the models used for basic scientific research in oncology mainly include tumor cell lines and human tumor xenograft models. Among them, the tumor cell line has the advantages of short culture cycle, easy operation, and low cost. The disadvantages are: 1) The tumor cell line is cultured under 2D conditions, which is quite different from the in vivo environment; 2) The genetics of the tumor cell line Background and gene expression profiles are quite different from clinical patient tumor cells. Therefore, tumor cell lines are usually only used as basic exploratory research tools. Human-derived xenograft model (patient-derived xenograft model, PDX model) refers to the ...

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
IPC IPC(8): C12N5/09C12Q1/02
CPCG01N33/5008C12N5/0693C12N2501/115C12N2501/415
Inventor 华国强湛先保李洁张丽星高韵程婉莹高登
Owner 上海赫佰生物医药科技有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap