Nucleus pulposus cell separation and in-vitro culture method

A technology for in vitro culture of nucleus pulposus cells, applied in cell dissociation methods, culture processes, tissue culture, etc., can solve problems such as difficulty in adhering to the wall, low cell survival rate, easy aging, degeneration, etc. And the effect of high tissue activity and easy adhesion

Inactive Publication Date: 2020-02-14
杭州东塘同年生物科技有限公司
View PDF4 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, there are various methods for the isolation and culture of nucleus pulposus cells, all of which have many disadvantages. The i

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
  • Nucleus pulposus cell separation and in-vitro culture method
  • Nucleus pulposus cell separation and in-vitro culture method
  • Nucleus pulposus cell separation and in-vitro culture method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0032] The present invention will be specifically introduced below in conjunction with specific embodiments.

[0033] A method for isolating and in vitro culturing nucleus pulposus cells, comprising the following contents:

[0034] The isolation of nucleus pulposus cells includes the following steps:

[0035] Step 1. After obtaining the nucleus pulposus tissue, place it in pre-cooled F12-DMEM culture medium containing 1% penicillin / streptomycin, transport it to the laboratory within 1 hour, and remove impurities; add 1% penicillin / streptomycin Streptomycin can prevent contamination during manipulation.

[0036] The specific process of removing impurities is as follows: wash with DPBS 2 to 3 times, centrifuge at 1500 rpm for 5 minutes, until no obvious blood stains; as a preferred method, DPBS is a DPBS mixed solution with penicillin / streptomycin added.

[0037] To obtain the nucleus pulposus tissue, the blood stains should be cleaned, and other tissues such as cartilage shou...

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 discloses a nucleus pulposus cell separation and in-vitro culture method. When nucleus pulposus cells are separated, nucleus pulposus tissues are obtained, and bloodstains and cartilagetissues need to be cleaned; the influence of miscellaneous cells on attachment and subsequent culture of nucleus pulposus cells is prevented, collagenase I is used as digestive juice, a digestion process is milder, the digested cells and tissues are higher in activity and easy to adhere to the wall, the digestion is terminated with a mixed solution containing fetal calf serum with the volume fraction of 20% and DPBS, the obtained cells are not prone to adhesion, bottle laying is more uniform, 1% of penicillin/streptomycin is added into a used reagent, and pollution to a sample or in collectionand transportation is prevented; during in-vitro culture, placenta mesenchymal stem cell culture supernatant freeze-dried powder reconstitution fluid is added, the adding amount is 10-40%, growth factors lacking in a culture medium are made up, and growth of nucleus pulposus cells is promoted; and the cultured cells are high in survival rate, easy to adhere to the wall, good in cell morphology and high and stable in quality.

Description

technical field [0001] The invention relates to the field of cell culture, in particular to a method for separating and in vitro culturing nucleus pulposus cells. Background technique [0002] Intervertebral disc degeneration (IVD) is the pathological basis of spinal degenerative diseases and one of the main causes of disc herniation and chronic low back pain. Neither conservative nor surgical treatment can fundamentally solve the problem of low back and leg pain caused by intervertebral disc degeneration. In recent years, emerging approaches for the treatment of degenerative intervertebral discs based on cells, especially stem cells, have attracted widespread interest. Studies have found that when the intervertebral disc degenerates, the degeneration of the nucleus pulposus tissue is the most obvious. Therefore, the related research of nucleus pulposus cells has become a hot spot. [0003] At present, there are various methods for the isolation and culture of nucleus pul...

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): C12N5/077
CPCC12N5/0669C12N2500/84C12N2509/00
Inventor 于元竹唐田娟刘芳
Owner 杭州东塘同年生物科技有限公司
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