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Human chondrocyte culture medium and application thereof

A chondrocyte and culture medium technology, applied in the field of cells, can solve the problems of limited scientific research, loss of chondrocyte function, etc., and achieve the effect of increasing the number of passages, promoting chondrocyte proliferation, and preventing transformation

Pending Publication Date: 2021-03-19
清远市人民医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In vitro expansion of monolayers leads to loss of chondrocyte function and a shift to a fibroblast-like phenotype; therefore, the properties of chondrocytes have limited their relevant scientific studies

Method used

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  • Human chondrocyte culture medium and application thereof
  • Human chondrocyte culture medium and application thereof
  • Human chondrocyte culture medium and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] A human chondrocyte culture medium, according to the volume fraction of each component, consists of 89% DMEM / F12, 10% fetal bovine serum, 20 μg / mL exosomes and 1% penicillin-streptomycin solution.

[0043] The preparation method of the medium is: take 87mL liquid DMEM / F12 liquid medium, 10mL fetal bovine serum, 2mL 1mg / mL exosomes and 1mL penicillin-streptomycin solution; Bovine serum, exosomes and penicillin-streptomycin solution. Adjust the pH of the medium to 7.0. During the specific preparation process, the volume of DMEM / F12 liquid medium was correspondingly reduced according to the amount (volume) of exosomes added.

Embodiment 2

[0045] A human chondrocyte culture medium, based on the volume fraction of each component, consists of 91.8% DMEM / F12, 7% fetal bovine serum, 22 μg / mL exosomes and 1.2% penicillin-streptomycin solution.

[0046] The preparation method of the medium is: take 89.6mL liquid DMEM / F12 liquid medium, 7mL fetal bovine serum, 2.2mL 1mg / mL exosomes and 1.2mL penicillin-streptomycin solution respectively; Add fetal bovine serum, exosomes and penicillin-streptomycin solution. Adjust the pH of the medium to 7.2. During the specific preparation process, the volume of DMEM / F12 liquid medium was correspondingly reduced according to the amount (volume) of exosomes added.

Embodiment 3

[0048] A human chondrocyte culture medium, based on the volume fraction of each component, consists of 85% DMEM / F12, 14% fetal bovine serum, 18 μg / mL exosomes and 1% penicillin-streptomycin solution.

[0049] The preparation method of the medium is: take 83.2mL liquid DMEM / F12 liquid medium, 14mL fetal bovine serum, 1.8mL 1mg / mL exosomes and 1mL penicillin-streptomycin solution respectively; Fetal bovine serum, exosomes, and penicillin-streptomycin solution were added. Adjust the pH of the medium to 7.0. During the specific preparation process, the volume of DMEM / F12 liquid medium was correspondingly reduced according to the amount (volume) of exosomes added.

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Abstract

The invention provides a human chondrocyte culture medium, and belongs to the technical field of cells. The invention optimizes a traditional culture medium, and by adding exosomes, fetalbovine serumand penicillin-streptomycin into a basal culture medium, chondrocyte proliferation can be effectively promoted, dedifferentiation of human chondrocytes is prevented, transformation of the human chondrocytes into fibrocyte-like phenotypes is prevented, and the single-layer culture passage number is increased. The invention also provides a method for isolating culture of the human chondrocytes by using the above-mentioned culture medium, and normal chondrocytes cultured to the sixth passage can be obtained.

Description

technical field [0001] The invention belongs to the field of cell technology, and in particular relates to a culture medium for human chondrocytes and an application thereof. Background technique [0002] Chondrocytes are an important type of cartilage cells. Various components of cartilage are basically differentiated or secreted by chondrocytes, including cartilage matrix and cartilage fibers. Chondrocytes can be cultured and expanded in vitro. Therefore, there are clinical cases of using chondrocytes to treat cartilage defects. [0003] Exosomes refer to small membrane vesicles containing proteins, lipids and nucleic acids, specifically vesicles with a diameter of 30-200 nm. Exosomes are easy to preserve, have stable biochemical activity, and have no potential toxicity. Human umbilical cord mesenchymal stem cells are widely used in clinical practice due to their advantages such as young cells, large amount of acquisition, no ethical issues, sufficient sources, high prol...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/077
CPCC12N5/0655C12N2500/84
Inventor 李鹏东姜文月齐玲赵贵芳张佳彭骞
Owner 清远市人民医院
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