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Mesenchymal stem cell induced differentiation and coating culture dish preparation

A technology of mesenchymal stem cells and culture dishes, applied in cell culture supports/coatings, animal cells, tissue culture, etc., can solve problems such as lack of biochemical and biomechanical properties, failure to restore cartilage, etc.

Inactive Publication Date: 2019-09-17
南京温博生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the above-mentioned techniques can maintain normal joint function and inhibit its further degeneration, they cannot restore damaged cartilage and do not have the same biochemical and biomechanical properties as native cartilage, requiring further development of regenerative medicine and permanent restoration of articular cartilage

Method used

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  • Mesenchymal stem cell induced differentiation and coating culture dish preparation
  • Mesenchymal stem cell induced differentiation and coating culture dish preparation
  • Mesenchymal stem cell induced differentiation and coating culture dish preparation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1: Xylan Directly Induces Osteogenic Differentiation of Umbilical Cord Mesenchymal Stem Cells

[0022] 1. Experimental materials

[0023] The umbilical cord tissue was taken from healthy pregnant women who underwent cesarean section, stored at 4°C, and the umbilical cord mesenchymal stem cells were isolated and cultured within 24 hours. The puerpera herself signed the informed consent form, stating that the umbilical cords taken could only be used for scientific research.

[0024] The cell incubator is a product of Thermo Fisher Scientific.

[0025] The flow cytometer is a product of BD Biosciences, USA.

[0026] Fetal bovine serum was the product of Gibco Company in the United States, α-MEM medium was the product of Hyclone Company, trypsin was purchased from Biyuntian, and chondrogenic differentiation medium was purchased from Guangzhou Bojin Biotechnology Co., Ltd.

[0027] Xylan was purchased from Shanghai Shifeng Biotechnology Co., Ltd. with a purity of ...

Embodiment 2

[0044] Embodiment 2: Preparation and application of xylan-coated culture dish

[0045] 1. Preparation of xylan-coated culture dishes

[0046] Preparation of xylan solution: Dissolve an appropriate amount of xylan in an aqueous solution of triethylamine with a pH value of 8.0, stir with a magnetic stirrer for 12 hours, adjust the pH value to neutral with glacial acetic acid, and prepare 10 g / L xylan Solution, store at 4°C and use within 8 hours.

[0047] Preparation of coated culture dish: take a sterile culture dish with a diameter of 100mm, add 50mL of the above xylan solution, dry at 50°C to form a film, rinse with deionized water several times, dry and store, a layer of film can be seen on the inner wall of the culture dish , the schematic diagram is as image 3 .

[0048] The coated Petri dishes were sterilized by ultraviolet irradiation and rinsed with culture medium before use.

[0049] 2. Induction and culture of umbilical cord mesenchymal stem cells in xylan-coated ...

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Abstract

The invention discloses mesenchymal stem cell induced differentiation and coating culture dish preparation. An agarose gel electrophoresis result shows that a culture group Aggrecan coated with no culture dish and type II collagen have no obvious expression, a group Aggrecan coated with a culture dish and type II collagen have obvious expression, it illustrates that the group coated with the culture dish has obvious expression of a cartilage marker, an xylan coated culture dish can be used for culturing human umbilical cord mesenchymal stem cells in vitro and inducing the mesenchymal stem cells into cartilage differentiation. An Aleian blue dyeing result shows that a control group is Aleian blue dying negative, the xylan group and the positive group are Aleian blue dyeing positive, it illustrates that the xylan group and the positive group both have obvious cartilage matrix protein glycan deposition, and it indicates that the xylan group and the positive group culture media can effectively induce human umbilical cord mesenchymal stem cells into cartilage differentiation.

Description

technical field [0001] The invention belongs to the biological field, and relates to the biological application of xylan inducing differentiation of mesenchymal stem cells and preparing coated culture dishes. Background technique [0002] Mesenchymal stem cells (MSCs) can differentiate into bone, cartilage, fat, muscle, endothelial and other tissue cells, and can even differentiate into retina, nerve cells, and hematopoietic cells across germ layers. They are important seed cells for cell therapy. [0003] Osteoarthritis is a rheumatic disease and one of the most common joint diseases in the world, with a prevalence of 10% in men and 18% in women over the age of 60. Osteoarthritis is a metabolic disease characterized by degeneration of articular cartilage, subchondral bone sclerosis, and synovitis, accompanied by bone friction, stiffness, pain, and limited mobility. Current treatments for cartilage repair focus on arthroscopic lavage, debridement, microfracture induction, a...

Claims

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Application Information

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IPC IPC(8): C12M1/22C12N5/0775C12N5/077
CPCC12M23/10C12M23/20C12N5/0655C12N2506/1369C12N2533/70
Inventor 姜东花张蕾
Owner 南京温博生物科技有限公司
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