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Endothelial cell culture medium and endothelial cell culture method

A technology of endothelial cells and culture medium, applied in the biological field, can solve the problem of poor proliferation ability of endothelial cells, and achieve the effect of improving the ability of angiogenesis, the ability of proliferation and vigorous growth.

Active Publication Date: 2015-10-14
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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Problems solved by technology

However, the proliferation ability of endothelial cells cultured in this medium is poor

Method used

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  • Endothelial cell culture medium and endothelial cell culture method
  • Endothelial cell culture medium and endothelial cell culture method
  • Endothelial cell culture medium and endothelial cell culture method

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preparation example Construction

[0051] In the present invention, the preparation method of the endothelial cell culture medium preferably includes the following steps:

[0052] Serum-free basal medium, insulin, transferrin, vitamin C, bovine serum albumin, basic fibroblast growth factor, vascular endothelial growth factor, stem cell growth factor, laminin, epidermal growth factor, non-essential amino acids, and metformin Hydrochloride was mixed to obtain endothelial cell culture medium.

[0053] In the present invention, the serum-free basal medium, insulin, transferrin, vitamin C, bovine serum albumin, basic fibroblast growth factor, vascular endothelial growth factor, stem cell growth factor, laminin, epidermal growth factor , the source and dosage of non-essential amino acids and metformin hydrochloride are the same as the serum-free basal medium, insulin, transferrin, vitamin C, bovine serum albumin, basic fibroblast growth factor, and vascular endothelial growth factor described in the above technical s...

Embodiment 1

[0081] 250 mg recombinant human insulin, 2.5 mg recombinant transferrin, 4.5 mg vitamin C, 5 g bovine serum albumin, 2.5 μg basic fibroblast growth factor, 5 μg vascular endothelial growth factor, 4 μg stem cell growth factor, 6 μg laminin, 4 μg Epidermal growth factor, 17.5 g of non-essential amino acids, 828.15 mg of metformin hydrochloride and 500 mL of IMDM basal medium were mixed to obtain endothelial cell culture medium.

[0082] Step 1: Dilute the umbilical cord blood with normal saline 1:1 and mix well;

[0083] Step 2: Take a 50mL centrifuge tube, add 15mL FICOLL lymphocyte separation medium, then add 25mL diluted umbilical cord blood, and centrifuge at 700g for 20 minutes;

[0084] Step 3: Aspirate the buffy coat layer, add 30mL normal saline to mix, centrifuge at 500g for 5 minutes, discard the supernatant, resuspend the pellet with the above-mentioned endothelial cell culture medium, and mix according to 5×10 5 Resuspend the cells at a density of / mL, add the cell...

Embodiment 2

[0102] 500 mg recombinant human insulin, 600 μg recombinant transferrin, 8 mg vitamin C, 5 g bovine serum albumin, 4 μg basic fibroblast growth factor, 10 μg vascular endothelial growth factor, 8.5 μg stem cell growth factor, 12 μg laminin, 5 μg epidermal growth factor Factors, 25g non-essential amino acids, 1407.855mg metformin hydrochloride and 500mL IMDM basal medium were mixed to obtain endothelial cell culture medium;

[0103] Step 1. Dilute the freshly collected umbilical cord blood with normal saline 1:1, and mix well;

[0104] Step 2: Take a 50mL centrifuge tube, add 15mL FICOLL lymphocyte separation medium, then add 25mL diluted umbilical cord blood, and centrifuge at 700g for 20 minutes;

[0105] Step 3: Absorb the buffy coat obtained in step 2, add 30mL of normal saline to mix, centrifuge at 500g for 5 minutes, discard the supernatant, resuspend the precipitate with medium, and mix according to 5×10 5 Resuspend the cells at a density of / mL, add the cell suspension...

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Abstract

The invention provides an endothelial cell culture medium and an endothelial cell culture method, wherein the endothelial cell culture medium comprises a serum-free basal culture medium, insulin, transferin, vitamin C, bovine serum albumin, a basic fibroblast growth factor, a blood vessel endothelium growth factor, a stem cell growth factor, laminin, an epidermal growth factor, non-essential amino acid and melbinum hydrochloride. By virtue of compatibility of ingredients, the endothelial cell culture medium provided by the invention is capable of improving the endothelial cell multiplication capacity. In addition, the endothelial cell culture medium provided by the invention is further capable of improving the blood vessel formation capability of endothelial cells. The experimental result indicates that when endothelial cells are cultured to the P10 generation by virtue of the endothelial cell culture medium provided by the invention, the cells still grow vigorously; the P10-generation endothelial cell phenotype CD309 and CD31 double-positive expression is as high as 97.2%.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a medium for endothelial cells and a method for culturing endothelial cells. Background technique [0002] Vascular endothelial cells, also known as endothelial cells, usually refer to the single layer of squamous epithelium that lines the inner surface of the heart, blood vessels, and lymphatic vessels, forming the inner walls of blood vessels. Endothelial cells are a general term for some phagocytes with common characteristics distributed in the brain, lymph nodes, lungs, liver, spleen and other organ tissues. They phagocytize foreign bodies, bacteria, necrotic and aging tissues, and also participate in collective immune activities, including: 1. Vasoconstriction and vasodilation to control blood pressure; 2. Coagulation (thrombosis and fibrinolysis); 3. Arteriosclerosis; 4. Angiogenesis; 5. Inflammation and swelling (such as: edema). 6. Endothelial cells also control some substan...

Claims

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Application Information

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IPC IPC(8): C12N5/071
Inventor 陈海佳王一飞葛啸虎万桦王小燕
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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