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

Osteosarcoma pulmonary metastasis primary cell strain and culture method and application thereof

A technology of primary cells and culture methods, applied in the field of primary cells of human osteosarcoma pulmonary metastases and their culture, which can solve the problems of small size, difficult separation, and no lung metastases

Active Publication Date: 2020-12-15
THE FIRST AFFILIATED HOSPITAL OF SUN YAT SEN UNIV
View PDF2 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the process of in vitro culture, there is a lack of microenvironment for the growth of the tumor in vivo, and after repeated passages, it adapts to the in vitro culture environment and changes, which cannot reflect the original characteristics of the tumor.
Pulmonary metastases are generally multiple in the lungs, and lesions appear in multiple locations, but the individual size is small, so it is difficult to obtain suitable specimens and difficult to separate after acquisition
Therefore, the existing osteosarcoma cell lines are all derived from in situ or distant metastases, and there are no cell lines with lung metastases.

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
  • Osteosarcoma pulmonary metastasis primary cell strain and culture method and application thereof
  • Osteosarcoma pulmonary metastasis primary cell strain and culture method and application thereof
  • Osteosarcoma pulmonary metastasis primary cell strain and culture method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1: Isolation and culture of osteosarcoma pulmonary metastases cell line

[0035] In a patient with osteosarcoma (on April 5, 2016, the right lower leg was amputated with osteosarcoma at the distal end of the tibia, and 14 times of adjuvant chemotherapy after surgery. CT of the posterior chest showed a nodule in the posterior basal segment of the left lower lobe, with a size of about 13.3x18.0mm. Thoracoscopic left lower lung wedge resection was performed on February 7, 2018.) The materials were collected on the day of the operation, and the non-necrotic area under the tumor capsule was excised during the operation (the specimen collection was approved by the Clinical Research and Experimental Animal Ethics Committee of the First Affiliated Hospital of Sun Yat-sen University ( Approval number: Lun Shen [2019] No. 060), the patient signed the informed consent). Viable samples from patients were washed with PBS by shaking to remove blood from the tissue. Cut the s...

Embodiment 2

[0039] Example 2: Detection of cell doubling time

[0040]Take 2000 ZOSL-1 cells, plant them in 96-well plates, add 100 μL culture medium (complete culture medium), add 10 μL cck-8 reagent the next day, incubate in the dark for 1 hour, detect the absorbance value at 450 nm, and replace with fresh culture medium liquid. Continuously detect for 7 days, draw the cell growth curve, and calculate the doubling time. Set up a blank control group, and add PBS solution to the peripheral wells to reduce evaporation.

[0041] The cell doubling time DT=t*[lg2 / (lgODt-lgOD0)] was calculated. Proliferation curve as figure 2 As shown, the doubling time is 39.28±3.04h.

Embodiment 3

[0042] Example 3: Detection of cell cycle and apoptosis

[0043] ZOSL-1 cells (P10, 90% full) were collected, and after washing, cell apoptosis was detected by flow cytometry AV / PI method. The result is as image 3 , 86.5% of the cells showed good activity.

[0044] ZOSL-1 cells (P10, 90% full) were collected, washed, fixed with ice ethanol, stained with endidine iodide, and detected cell cycle by flow cytometry. The result is as Figure 4 , 50.77% in G1 phase, 15.37% in G2 phase, and 33.85% in S phase.

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 provides a human osteosarcoma pulmonary metastasis primary cell strain and a culture method and application thereof. The human osteosarcoma pulmonary metastasis primary cell strain is named as ZOSL-1, and the preservation number is CCTCC No: C202088. The human osteosarcoma pulmonary metastasis primary cell strain is obtained by combining a pancreatin differential digestion method, adifferential adherence method and a serum-free medium stimulation method and screening purified tumor cells, can be used for osteosarcoma pulmonary metastasis mechanism research and antitumor drug research and has good medical value.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to primary cells of human osteosarcoma pulmonary metastases and a culture method and application thereof. Background technique [0002] Osteosarcoma (OS), also known as osteosarcoma, is more common in adolescents and children. Lung metastasis is the leading cause of death in patients with osteosarcoma. Appropriate preclinical models are crucial for obtaining more effective treatment options. [0003] Cell line is one of the commonly used preclinical models, which has the advantages of convenience and speed. However, in the process of in vitro culture, there is a lack of microenvironment for the growth of the tumor in vivo, and after repeated passages, it adapts to the in vitro culture environment and changes, which cannot reflect the original characteristics of the tumor. Pulmonary metastases are generally multiple in the lungs, and lesions appear in multiple locations, but the indiv...

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): C12N5/09C12N5/077C12Q1/02C12R1/91
CPCC12N5/0693C12N5/0654G01N33/5011C12N2509/00C12N2509/10G01N2500/10
Inventor 尹军强沈靖南赵巨鹏樊泽培
Owner THE FIRST AFFILIATED HOSPITAL OF SUN YAT SEN UNIV
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