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Endothelial cell culture solution

An endothelial cell and culture medium technology, applied in cell culture medium, vascular endothelial cells, cell culture active agents, etc., can solve the problems of increasing the cost of culture medium and not being conducive to the stability control of culture medium batches.

Active Publication Date: 2017-02-22
沈阳细胞治疗工程技术研发中心有限公司
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Problems solved by technology

These substances may carry unknown pathogens, including pathogenic prion proteins and foreign proteins targeting fetal bovine antigens, which may cause allergic reactions after being injected into the human body; in addition, in order to make endothelial cells grow faster and better, patent 201310025943.5 The addition of a large number of growth factors that promote the growth of endothelial cells in the in vitro culture expansion system not only increases the cost of the culture medium, but also the introduction of multiple growth factors is not conducive to the stability control of the culture medium batches

Method used

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Embodiment 1

[0027] 1. The preparation method of endothelial cell culture medium, the specific steps are as follows.

[0028] Step 1. Prepare human platelet lysate: take 2×10 11 L -1 Add human platelets collected by machine into a 250ml centrifuge bottle, and centrifuge at 6000g for 20 minutes; remove the supernatant, add normal saline to wash the platelets, and suspend them with 15ml of normal saline after centrifugation; freeze overnight in a -20°C refrigerator for 12 hours, and then place at room temperature Melt down; carry out three cycles in this way to obtain platelet lysate.

[0029] Step 2: Centrifuge 100ml of human platelet lysate, 60mg of penicillin, 100mg of streptomycin, 292mg of L-glutamine, 50mg of vitamin C, 30mg of laminin 10, 30mg of laminin 11 and 40mg of fibronectin After mixing, the basal medium DMEM was added to a volume of 1 L; the pH value was 6.8-7.0, and the osmotic pressure was 310 mOsm / kg to obtain endothelial cell culture medium.

[0030] 2. The endothelial ...

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Abstract

The invention belongs to the technical field of biology, and particularly relates to endothelial cell culture solution. The endothelial cell culture solution comprises a basic culture medium DMEM (dulbecco modified eagle medium), human platelet lysate, L-glutamine, vitamin C, laminin 10, laminin 11, fibronectin, penicillin and streptomycin. The endothelial cell culture solution has the advantages that a pH (potential of hydrogen) value of the endothelial cell culture solution is 6.8-7.0, and the osmotic pressure of the endothelial cell culture solution is 310 mOsm / kg; endothelial cells cultured by the endothelial cell culture solution are high in adherence and proliferation speed, purity, migration capacity and lumen formation rate, invasion effects of the endothelial cells can be inhibited, and accordingly the endothelial cell culture solution is a perfect endothelial cell culture system.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to an endothelial cell culture solution. Background technique [0002] Endothelial cells are a thin layer of epithelial cells that have the functions of selective barrier, hemostasis, anticoagulation and transport, and can participate in the body's inflammatory response. Studies have shown that endothelial cells are closely related to the occurrence and development of atherosclerosis, and various active factors secreted by endothelial cells participate in the body's response and maintain the stable state of life. [0003] In recent years, the research on endothelial cells has been increasing day by day, and the research results are gradually moving towards clinical application, among which the in vitro expansion and culture of endothelial cells is an important part of the research and application process. However, under normal conditions, endothelial cells have poor proliferative abili...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
CPCC12N5/0037C12N5/069C12N2500/32C12N2500/38C12N2500/84C12N2500/90C12N2501/998
Inventor 杨柳松
Owner 沈阳细胞治疗工程技术研发中心有限公司
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