hCMEC/D3〖Immortalized human brain microvascular endothelial cells〗

/〖Immortalized human brain microvascular endothelial cells〗
Cell number
Cell abbreviation
/cell
Cell name
/(Immortalized human brain microvascular endothelial cells)
Detailed background
See cell instructions
Source of cells
Main source of cells and establishment of departments of a few well-known universities at home and abroad
Precautions for purchasing cells
. After receiving the cells, first Observe whether the cell bottle is intact and whether the culture medium is leaking or turbid. If the above phenomena occur, please contact us in time.
. Read the cell instructions carefully to understand the cell-related information, such as cell morphology, culture medium used, serum ratio, required cytokines, etc.
. Wipe the surface of the cell bottle with % alcohol and observe the cell status under a microscope. /〖Immortalized human brain microvascular endothelial cells〗
Due to transportation problems, a small amount of adherent cells will fall off the bottle wall. Place the cells in an incubator and culture overnight, and then take them out for observation the next day. At this time, most cells will adhere to the wall. If the cells still cannot adhere, please use trypan blue staining to determine the cell viability. If the cell viability is confirmed to be normal, please centrifuge the cells and culture them again with fresh culture medium. If the staining result shows that the cells have no cells. Vitality, please take a photo and contact us in time. After the information is confirmed, we will send it to you for free again.
. It is recommended that customers take a few photos of the cells the day before and after receiving them to record the status of the cells to facilitate communication with the company's technical department.
Packaging specifications
Resuscitation culture bottle (one bottle) or cryopreservation vials (two)
Cell specifications
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Safety level
Cell species
Human
Tissue source
Brain tissue blood-brain barrier
Cell morphology
Endothelium
Cell characteristics
Adhesion
Cell fusion degree
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Cell detection
Cells do not contain, mycoplasma, bacteria, yeast and fungi
Three Don’ts for cell culture
Raising scientific research cells is for biomedical research Basic skills. I have three little experiences to share with you
. Don’t flask the mouth of the bottle. Most people prefer to close the bottle over the flame of an alcohol burner, feeling that this avoids contamination. I wonder if you are confused about why the culture medium becomes more purple the more you use it? Do you know why? The mouth of the flask is burned. The culture medium generally uses carbonic acid/bicarbonate as a buffer system. When the mouth of the bottle is over the flame, hot air enters the bottle. After tightening the stopper and placing it in the refrigerator, the air becomes cold and the pressure drops suddenly, and the carbonic acid in the culture medium is converted into carbon dioxide and released. If there is less carbonic acid in the culture medium, it will of course become alkaline. If there is no flask mouth, you will find that the rate at which the culture solution becomes alkaline will be greatly slowed down. (Of course, it will still become more or less alkaline with more use, because the room temperature is still higher than the temperature in the refrigerator)
In fact, the mouth of the flask to prevent contamination is purely a psychological comfort. You might as well try shaking your fingers over the fire a few times, and you will find that there is no burning pain at all. If there are bacteria, shaking them twice won't kill them much. If you want to burn all the bacteria, you have to burn the bottle mouth and stopper red. I have never used flasks in the country. After coming to the United States, I found that this place is more thorough. There is no alcohol lamp at all in the ultra-clean bench.
. Do not look at cells frequently. For beginners, raising cells is like raising a child, so you should check it out when you have time. The worse the cells are maintained, the more often they need to be seen. In fact, looking at the cells does not bring any benefit to the growth of the cells, especially for primary cells or cells with particularly low density after immunomagnetic bead sorting. The growth factors in the culture medium are very limited. The cells finally secrete some growth-promoting factors around them, forming a local environment conducive to growth. You ran to look at the cells, and the culture bottle shook like this, shaking all the factors away. It is often seen that novices are looking at cells all day long, but the cells never grow well. The old hand throws away the cells for several days and the cells grow like crazy.
. Don’t be afraid of digestion. During passaging, beginners are often afraid of excessive cell digestion and will terminate digestion early. Before the cells came down, I blew the lamp so hard that the bottle was filled with bubbles. In my opinion, vigorous pipetting causes more damage to the cells than digestion. When my senior brother was doing primary vascular smooth muscle, he digested it overnight and the cells were fine. I usually wait until the cells become completely round before stopping digestion. The cells can be removed with a gentle shake. Also, be gentle and try not to create bubbles. The mechanical stress when the bubble bursts is quite large, and the damage to cells is also great.
See the cell instructions for details
Passaging method
It is recommended to change the medium every two days
Freezing conditions
See the cell instructions for details
Main literature
Please Please refer to relevant published articles for details