Human synovial fluid mesenchymal stem cell culture medium and preparation method thereof

A technology of mesenchymal stem cells and joint fluid, which is applied in the field of culture medium and its preparation for human joint fluid mesenchymal stem cells, which can solve the problems of slow growth, poor medium culture effect, poor quality of single cell images and fluorescent staining photos Good and other issues, to achieve the effect of good promotion value

Pending Publication Date: 2018-11-23
THE SECOND PEOPLES HOSPITAL OF SHENZHEN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In the prior art, the culture medium for human synovial fluid mesenchymal stem cells is not effective, the growth rate is slow, and the quality of single cell images and fluorescent staining photos of mesenchymal stem cells isolated by image flow cytometry is poor. Good; therefore, the present invention proposes a culture medium for human synovial fluid mesenchymal stem cells and a preparation method thereof

Method used

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Embodiment 1

[0020] The invention provides a technical solution: a culture medium for human synovial fluid mesenchymal stem cells, the formula of the medium for human synovial fluid mesenchymal stem cells includes the following components: fetal bovine serum 9%, penicillin 12% , streptomycin 4%, amino acid 2%, glucose 3%, anhydrous calcium chloride 4%, leucine 1%, potassium chloride 5%, hypoxanthine 1%, thymidine 2%, riboflavin 3 %, phenylalanine 2%, fluorescent antibody 3%, and the rest is water.

[0021] Among them, the fluorescent antibody is PE-CD44.

[0022] A method for preparing a culture medium for human synovial fluid mesenchymal stem cells as described above, the preparation method comprising the following steps:

[0023] S1: Quantitative weighing, all raw materials are weighed according to the above components;

[0024] S2: Pour into the culture medium, pour all amino acids, glucose, anhydrous calcium chloride, leucine, potassium chloride, hypoxanthine, thymidine, riboflavin, ...

Embodiment 2

[0029] The invention provides a technical solution: a medium for human synovial fluid mesenchymal stem cells, the formula of the medium for human synovial fluid mesenchymal stem cells includes the following components: 10% fetal bovine serum, penicillin 13 %, streptomycin 5%, amino acid 3%, glucose 4%, anhydrous calcium chloride 5%, leucine 2%, potassium chloride 6%, hypoxanthine 2%, thymidine 3%, riboflavin 4%, phenylalanine 3%, fluorescent antibody 4%, and the rest is water.

[0030] Among them, APC-CD73 is selected as the fluorescent antibody.

[0031] A method for preparing a culture medium for human synovial fluid mesenchymal stem cells as described above, the preparation method comprising the following steps:

[0032] S1: Quantitative weighing, all raw materials are weighed according to the above components;

[0033] S2: Pour into the culture medium, pour all amino acids, glucose, anhydrous calcium chloride, leucine, potassium chloride, hypoxanthine, thymidine, ribofla...

Embodiment 3

[0038] The invention provides a technical solution: a medium for human synovial fluid mesenchymal stem cells, the formula of the medium for human synovial fluid mesenchymal stem cells includes the following components: fetal bovine serum 11%, penicillin 14 %, streptomycin 6%, amino acid 4%, glucose 5%, anhydrous calcium chloride 6%, leucine 3%, potassium chloride 7%, hypoxanthine 3%, thymidine 4%, riboflavin 5%, phenylalanine 4%, fluorescent antibody 5%, and the rest is water.

[0039] Among them, FITV-CD90 was selected as the fluorescent antibody.

[0040] A method for preparing a culture medium for human synovial fluid mesenchymal stem cells as described above, the preparation method comprising the following steps:

[0041] S1: Quantitative weighing, all raw materials are weighed according to the above components;

[0042] S2: Pour into the culture medium, pour all amino acids, glucose, anhydrous calcium chloride, leucine, potassium chloride, hypoxanthine, thymidine, ribof...

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Abstract

The invention discloses a human synovial fluid mesenchymal stem cell culture medium. A formula of the human synovial fluid mesenchymal stem cell culture medium comprises the following ingredients: 9%to 11% of fetal calf serum, 12% to 15% of penicillin, 4% to 7% of streptomycin, 2% to 6% of amino acid, 3% to 5% of glucose, 4% to 6% of anhydrous calcium chloride, 1% to 4% of leucine, 5% to 9% of potassium chloride, 1% to 3% of hypoxanthine, 2% to 4% of thymidine, 3% to 5% of riboflavin, 2% to 5% of phenylalanine, 3% to 8% of fluorescence antibody and the balance of water. The invention furtherdiscloses a preparation method of the human synovial fluid mesenchymal stem cell culture medium. The preparation method comprises the following steps: S1, quantitatively weighing; S2, pouring the culture medium; S3, cleaning stem cells; S4, observing and culturing; S5, dyeing and marking. By means of the prepared human synovial fluid mesenchymal stem cell culture medium disclosed by the invention,the stem cells are visually in a spindle shape under a microscope, colony is formed, and the appearance characteristic of the synovial fluid mesenchymal stem cells can be met.

Description

technical field [0001] The invention relates to the technical field of medium preparation, in particular to a medium for human synovial fluid mesenchymal stem cells and a preparation method thereof. Background technique [0002] Osteoarthritis is one of the most common joint diseases. Its prominent case feature is progressive destruction of articular cartilage, which leads to joint pain and limited mobility, which seriously affects the quality of life of patients. Mesenchymal stem cells are derived from mesenchymal Tissues, cells with proliferation and multi-directional differentiation capabilities, are widely involved in the process of injury repair in vivo. Previous studies have found that mesenchymal stem cells are contained in the joint fluid, and the number of mesenchymal stem cells in the joint fluid of early osteoarthritis is higher than that of normal people ; [0003] In the prior art, the culture medium for human synovial fluid mesenchymal stem cells is not effect...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
CPCC12N5/0668C12N2500/14C12N2500/12C12N2500/38C12N2500/34C12N2500/40C12N2500/32C12N2501/50C12N2501/599
Inventor 王大平段莉李兴福徐晓熊建义欧阳侃朱飞燕刘威黄江鸿邓志钦蒯声政
Owner THE SECOND PEOPLES HOSPITAL OF SHENZHEN
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