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Primary culture method of elderly rat brain vascular endothelial cell

A cerebrovascular endothelium and culture method technology, applied in the primary culture of cerebral vascular endothelial cells of aged rats, in the biological field, can solve the problem that the sample objects and culture techniques cannot meet the aged blood-brain barrier model, consume a long time, and cannot be frozen, etc. problem, to save time and money, save time and money, increase purity

Inactive Publication Date: 2013-04-24
SHANDONG UNIV
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  • Abstract
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AI Technical Summary

Problems solved by technology

[0002] The existing primary culture of rat cerebrovascular endothelial cells is obtained from neonatal rats (<10 days, 35-50g), juvenile rats (<3weeks, 80-100g) or young adult rats (2-3 months old) (200g- 300g), and the cultured cerebrovascular endothelial cells can only be passaged once and cannot be frozen. Using the primary cultured rat cerebrovascular endothelial cells as experimental materials leads to the disadvantages of long time consumption and high cost, such as: Bowman PD, Betz AL, Ar D, Wolinsky JS, Penney JB, Shivers RR, Goldstein GW.1981.Primary Culture of Capillary Endothelium From Rat Brain.IN VITRO.17(4),353-362
[0003] The precedent of successful culture of cerebrovascular endothelial cells in aged rats (19-22 months, 900g-1200g) has not been reported so far; the existing materials and culture techniques cannot meet the requirements of establishing the aged blood-brain barrier model in vitro and studying the effects of the aged blood-brain barrier. Physiological properties, and requirements for blood-brain barrier-related degeneration in geriatric neurological diseases

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  • Primary culture method of elderly rat brain vascular endothelial cell
  • Primary culture method of elderly rat brain vascular endothelial cell
  • Primary culture method of elderly rat brain vascular endothelial cell

Examples

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Effect test

Embodiment 1

[0070] A method for primary culture of aged rat cerebrovascular endothelial cells, the steps are as follows:

[0071] (1) Decapitate a 20-month-old aged rat, peel off the skin of the rat’s head, and then soak the rat’s head in a separation solution containing 2% (volume percent) fetal bovine serum (FBS) at 0°C for 5 Minutes, take it out, cut the two cerebral hemispheres, roll the two cerebral hemispheres on sterilized ordinary filter paper for a week, then remove the brainstem and white matter, and keep the gray matter to obtain the sample tissue;

[0072] The separation solution is based on the DMEM medium, and the following components are added per 100 ml: 1 ml penicillin-streptomycin solution; 100 μl thiopurine gentamycin;

[0073] (2) Place the sample tissue in a 5ml ice-cold Petri dish containing 8% (volume percent) FBS separation solution and cut it into 1-2mm 3 Add 8% (volume percent) FBS separation solution to 25ml of small sample tissue, and disperse by pipetting, th...

Embodiment 2

[0082] A method for primary culture of aged rat cerebrovascular endothelial cells, the steps are as follows:

[0083] (1) Decapitate 22-month-old aged rats, peel off the skin of the rat's head, and then soak the rat's head in a separation solution containing 2% (volume percent) fetal bovine serum (FBS) at 0°C for 10 Minutes, take it out, cut the two cerebral hemispheres, roll the two cerebral hemispheres on sterilized ordinary filter paper for a week, then remove the brainstem and white matter, and keep the gray matter to obtain the sample tissue;

[0084] The separation solution is based on the DMEM medium, and the following components are added per 100 ml: 1 ml penicillin-streptomycin solution; 100 μl thiopurine gentamycin;

[0085] (2) Place the sample tissue in a 5ml ice-cold Petri dish containing 8% (volume percent) FBS separation solution and cut it into 1-2mm 3 Add 8% (volume percent) FBS separation solution to 25ml of small sample tissue, and disperse by pipetting, th...

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Abstract

The invention relates to a primary culture method of elderly rat brain vascular endothelial cells. The primary culture method comprises the following steps of firstly, taking out a grey matter part of an elderly rat brain tissue, and making a sample tissue; secondly, putting into a culture dish with a separating medium of fetal calf serum, and carrying out blowing, dispersing and biological enzyme digestion so as to prepare dispersed cells; thirdly, centrifuging so as to prepare a blood capillary; fourthly, re-suspending the blood capillary in the separation medium, and digesting to prepare blood capillary suspension; fifthly, centrifuging, re-suspending the sediment in the separation medium, and carrying out Percoll concentration gradient centrifugation so as to prepare the brain vascular endothelial cells; and sixthly, putting in a culture medium, culturing and carrying out digestion passage or freezing and storing. The primary culture method adopts low-concentration pancreatin or PBS (Phosphate Buffer Solution) digestion cells with 1m M EDTA (Ethylene Diamine Tetraacetic Acid), the cells can be subjected to three times of digestion passage but the characteristics of the brain vascular endothelial cells are still maintained, and the purity and the characteristics of the brain vascular endothelial cells are still maintained after being subjected to freezing and recovery, as a result, a great deal of time and money are saved.

Description

technical field [0001] The invention relates to a primary culture method for aged rat cerebral vascular endothelial cells, belonging to the technical field of biotechnology. Background technique [0002] The existing primary culture of rat cerebrovascular endothelial cells is obtained from neonatal rats (<10 days, 35-50g), juvenile rats (<3weeks, 80-100g) or young adult rats (2-3 months old) (200g- 300g), and the cultured cerebrovascular endothelial cells can only be passaged once and cannot be frozen. Using the primary cultured rat cerebrovascular endothelial cells as experimental materials leads to the disadvantages of long time consumption and high cost, such as: Bowman PD, Betz AL, Ar D, Wolinsky JS, Penney JB, Shivers RR, Goldstein GW. 1981. Primary Culture of Capillary Endothelium From Rat Brain. IN VITRO. 17(4), 353-362. [0003] The precedent of successful culture of cerebrovascular endothelial cells in aged rats (19-22 months, 900g-1200g) has not been reporte...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
Inventor 王丽梅崔敏王越陈哲宇
Owner SHANDONG UNIV
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