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Application of progesterone in stimulation of fibronectin secretion of mesenchymal stem cells

A mesenchymal stem cell and protein technology, which is applied in the application field of progesterone to stimulate mesenchymal stem cells to secrete fibronectin, can solve the problems of high price and reduce the amount of exogenous addition, and achieve the effect of reducing costs

Inactive Publication Date: 2015-10-28
河南中科干细胞基因工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0018] For the existing serum-free medium for culturing mesenchymal stem cells, it is necessary to add substances that promote cell adhesion, such as fibronectin, vitronectin, collagen and other substances, and the prices of fibronectin and other substances are very expensive. The purpose of the present invention When preparing serum-free medium for mesenchymal stem cells, add a low-cost small molecule substance, use the activity of the cell itself, use this substance to stimulate the function of the cell to secrete adhesion molecules, and stimulate the secretion of the cell through the low-cost small molecule substance Fibronectin, which replaces the expensive fibronectin that must be added, thereby reducing the amount of exogenous additions, without affecting cell proliferation. After culture and passage, mesenchymal stem cells still express characteristic surface molecules to protect mesenchymal stem cells. adherent growth

Method used

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  • Application of progesterone in stimulation of fibronectin secretion of mesenchymal stem cells
  • Application of progesterone in stimulation of fibronectin secretion of mesenchymal stem cells
  • Application of progesterone in stimulation of fibronectin secretion of mesenchymal stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Observation of cell morphology

[0035] 1. Experimental materials

[0036] (1) Cells: human bone marrow mesenchymal stem cells, third generation, derived from healthy people, cryopreserved by our company.

[0037] (2) Progesterone: It is a product of Sigma Company in the United States, and the article number is P0130. The weighed powder was dissolved in DMSO, and stored in aliquots at 4°C.

[0038] (3) ITS: It is a compound preparation, used for serum substitutes, and it is a product of Sigma Company. The final concentration contains 10 μg / ml insulin, 5.5 μg / ml transferrin, 5 ng / ml sodium selenite, 4.70 μg / ml linoleic acid, 4.70 μg / ml linolenic acid, and 500 μg / ml bovine serum albumin.

[0039] The stock solution provided by Cat. No. I2771.Sigma is 100X.

[0040] (4) Alpha modified minimum essential medium (alpha minimum essential medium, alpha-MEM) is a basic medium commonly used for culturing mesenchymal stem cells.

[0041] It is a product of Sigma Company, the ...

Embodiment 2

[0049] Western blot hybridization

[0050] 1. Test material

[0051] (1) Cells: 2 strains of human bone marrow mesenchymal stem cells, the third generation, respectively derived from two healthy people, cryopreserved by our company.

[0052] (2) Progesterone: It is a product of Sigma Company in the United States, the article number is P0130; the weighed powder is dissolved in DMSO, and stored in separate packages at 4°C.

[0053] (3) ITS: It is a compound preparation, used for serum substitutes, and it is a product of Sigma Company. The final concentration contains 10 μg / ml insulin, 5.5 μg / ml transferrin, 5 ng / ml sodium selenite, 4.70 μg / ml linoleic acid, 4.70 μg / ml linolenic acid, and 500 μg / ml bovine serum albumin. The stock solution provided by Cat. No. I2771.Sigma is 100X.

[0054] (4) Alpha modified minimum essential medium (alpha minimum essential medium, alpha-MEM) is a basic medium commonly used for culturing mesenchymal stem cells. It is a product of Sigma Company...

Embodiment 3

[0063] Cell Adhesion Assay

[0064] 1. Materials and Reagents

[0065] (1) Cells: 3 strains of human bone marrow mesenchymal stem cells, the third generation, were derived from three healthy people, and were cryopreserved by our company.

[0066] (2) Progesterone: It is a product of Sigma Company in the United States, and the article number is P0130. The weighed powder was dissolved in DMSO, and stored in aliquots at 4°C.

[0067] (3) ITS: It is a compound preparation, used for serum substitutes, and it is a product of Sigma Company. The final concentration contains 10 μg / ml insulin, 5.5 μg / ml transferrin, 5 ng / ml sodium selenite, 4.70 μg / ml linoleic acid, 4.70 μg / ml linolenic acid, and 500 μg / ml bovine serum albumin. The stock solution provided by Cat. No. I2771.Sigma is 100X.

[0068] (4) Alpha modified minimum essential medium (alpha minimum essential medium, alpha-MEM) is a basic medium commonly used for culturing mesenchymal stem cells. It is a product of Sigma Compa...

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Abstract

The invention belongs to the field and tissue culture, and especially relates to an application of progesterone in stimulation of the fibronectin secretion of mesenchymal stem cells. Fibronectins are needed when mesenchymal stem cells are cultured under a serum-free or low-serum condition to increase the adhesion of the mesenchymal stem cells, and the fibronectins are expensive, so the culture cost is high. The invention discloses a micro-molecular substance progesterone also called luteosterone or pregnenedione. The molecular formula of progesterone is C21H30O2, the molecular weight is 314.46, and progesterone can stimulate mesenchymal stem cells to synthesize the fibronectins in order to promote adhesion of the mesenchymal stem cells. Adhesion molecules needed by the cells are synthesized by the cells through processing the cells by using progesterone via utilizing the activity of the mesenchymal stem cells, so the cost of the serum-free or low-serum culture medium of the mesenchymal stem cells is greatly reduced, the adhesion ability of the progesterone processed mesenchymal stem cells is enhanced, and the progesterone processed mesenchymal stem cells can be used for constructing engineered tissues based on the mesenchymal stem cells.

Description

technical field [0001] The invention belongs to the field of cell and tissue culture, in particular to the application of progesterone to stimulate mesenchymal stem cells to secrete fibronectin. Background technique [0002] Progesterone (progesterone) Progesterone is a natural progesterone secreted by the corpus luteum of the ovary. It has a significant morphological effect on the endometrium stimulated by estrogen in vivo and is necessary for maintaining pregnancy. [0003] The main functions of progesterone are: [0004] 1. Prepare for the implantation of fertilized eggs: This is one of the main functions of progesterone. In the late menstrual cycle of women, progesterone will make the glands in the uterine mucosa grow, the uterus will be congested, the endometrium will thicken, and it will be fertilized eggs. , get ready; after the implantation of the fertilized egg, it will produce the placenta, and it will also reduce the excitability of the pregnant uterus, inhibit i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/02C12N5/0775
Inventor 郭子宽赵靖王娟李乔丽芦俊萍刘瑾尹珊
Owner 河南中科干细胞基因工程有限公司
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