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Method for constructing soft tissues of mesenchymal stem cells of synovial membranes and application of method

A technology of mesenchymal stem cells and construction methods, applied in the field of soft tissue construction of synovial mesenchymal stem cells, to achieve good application prospects, easy to obtain materials, and good cartilage differentiation ability

Active Publication Date: 2018-12-07
SHANGHAI FIRST PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But a kind of reagent for repairing cartilage damage of the present invention and application thereof have not yet been reported

Method used

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  • Method for constructing soft tissues of mesenchymal stem cells of synovial membranes and application of method
  • Method for constructing soft tissues of mesenchymal stem cells of synovial membranes and application of method
  • Method for constructing soft tissues of mesenchymal stem cells of synovial membranes and application of method

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] The preparation method of embodiment 1 soft tissue (1)

[0036] The soft tissue was prepared as follows:

[0037](1) Isolation and culture of synovial mesenchymal stem cells: The fresh human synovial specimens were surface-sterilized twice with 75% ethanol, washed twice with PBS, placed in a petri dish, and the tissues were cut with sterilized surgical scissors. For small fragments, put them in a 50ml centrifuge tube, add an appropriate amount of PBS, centrifuge, discard the supernatant, add an appropriate amount of type II collagenase, shake on a shaker (200rpm, 37°C) for 2 hours, centrifuge to discard the collagenase, and finally in the tissue Add an appropriate amount of low-sugar DMEM culture solution containing 10% FBS to the fragments, then transfer to a petri dish and place at 37°C, 5% CO 2 Culture in an incubator with saturated humidity, transfer the tissue block to a new culture dish two days later, continue culturing for two days and then discard the tissue b...

Embodiment 2

[0042] Embodiment 2 The preparation method of soft tissue (two)

[0043] The soft tissue was prepared as follows:

[0044] (1) Isolation and culture of synovial mesenchymal stem cells: sterilize the surface of fresh human synovial specimens with 70% ethanol once, wash them once with PBS, put them in a petri dish, and cut the tissue with sterilized surgical scissors. For small fragments, put in a 50ml centrifuge tube, add appropriate amount of PBS, centrifuge, discard the supernatant, add appropriate amount of type II collagenase, shake on a shaker (200rpm, 37°C) for 1 hour, centrifuge to discard the collagenase, and finally in the tissue Add an appropriate amount of low-sugar DMEM medium containing 5% FBS to the fragments, then transfer to a petri dish, and place at 32°C, 3% CO 2 Culture in an incubator with saturated humidity, transfer the tissue block to a new culture dish two days later, continue to culture for two days and then discard the tissue block until the mesenchym...

Embodiment 3

[0049] Embodiment 3 The preparation method of soft tissue (three)

[0050] The soft tissue was prepared as follows:

[0051] (1) Isolation and culture of synovial mesenchymal stem cells: The fresh human synovial specimens were surface-sterilized with 80% ethanol for 3 times, washed with PBS for 3 times, placed in a petri dish, and the tissues were cut with sterilized surgical scissors. For small fragments, put in a 50ml centrifuge tube, add appropriate amount of PBS, centrifuge, discard the supernatant, add appropriate amount of type II collagenase, shake on a shaker (200rpm, 37°C) for 3 hours, centrifuge to discard the collagenase, and finally in the tissue Add an appropriate amount of low-sugar DMEM culture solution containing 15% FBS to the fragments, then transfer to a petri dish and place at 42°C, 7% CO 2 Culture in an incubator with saturated humidity, transfer the tissue block to a new culture dish two days later, continue to culture for two days and then discard the t...

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Abstract

The invention relates to a method for constructing soft tissues of mesenchymal stem cells of synovial membranes. According to the method, the soft tissues are simply and effectively manufactured through four steps of separating and culturing the mesenchymal stem cells of the synovial membranes, authenticating the mesenchymal stem cells of the synovial membranes, culturing and separating the soft tissues and detecting cartilage differentiation capacity, and the differentiation capacity of cartilages is detected, so that cartilages are effectively repaired. The method has the advantages that (1)the constructed soft tissues have good cartilage differentiation capacity, and a new choice is provided for the repairing of the cartilages; and (2) the sampling is convenient, the detection is easy,and the method has good application prospects.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a soft tissue construction method and application of synovial mesenchymal stem cells. Background technique [0002] Articular cartilage defect is a relatively common clinical disease, mainly manifested as intractable pain and progressive joint dysfunction. If it is not cured in time, it will eventually lead to degenerative osteoarthritis. With the aging process of the whole country and even the world, such diseases will undoubtedly bring long-term and heavy burdens to families and society. Articular cartilage is a layer of dense connective tissue on the surface of the joint, mainly composed of cartilage matrix and chondrocytes, lacking blood vessels, lymph and nerve tissue, so the metabolic activity of chondrocytes is low. In addition, although chondrocytes have a certain ability to regenerate, the number of chondrocytes is very small (accounting for only about 5% of cartila...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
CPCC12N5/0668C12N2500/30C12N2501/50C12N2501/599C12N2501/734C12N2509/00
Inventor 马春辉王刚吕琦易诚青腾松松
Owner SHANGHAI FIRST PEOPLES HOSPITAL
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