Diffuse large B cells h:r
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[Product number] h:r
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[Chinese abbreviation] h:r
cell
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[Chinese name] h:r
(diffuse large B cell)
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[Background information] h:r
See cells Instructions
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[Product source] h:r
Main source of cells, DMZ,, R, rm,,,, B, rd, and a few well-known universities at home and abroad
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The company provides cell characteristics such as adherent, semi-adherent, suspended, semi-suspended, adherent and suspended mixture. Generally, the B volume of cell culture is %. If the cell status is poor, it can be increased. The amount of B reaches %. After the cells are stable, adjust the amount of B back to %. For first-time experimenters, in general cell culture, double antibodies need to be added to prevent cell contamination. When the cell confluence reaches %, we start sending diffuse large B cells< br/>This can maintain good cell viability when cells are received. Precautions for resuscitating cells
Change the medium on the same day after receiving the cells. Replace all cells with the customer's own fresh culture medium, put them in the incubator, and digest them the next day.
Observe while digesting. According to the shape of the cells, stop the digestion time and pipe the edges of the cells at half-minute intervals. If the cells can be piped down, the digestion will be terminated. Customers explore for better digestion time.
There is a cell operation manual accompanying the cells, please read it carefully.
Remember to add % double resistance to reduce the probability of contamination. In addition, freeze the seeds as soon as possible for later use.
The after-sales time is one week, which is limited to quality problems of the cells themselves. Cell problems caused by Party B's own improper operations (irregular operation, over-digestion, contamination caused by not adding double-antibodies, etc.) are not included in the after-sales scope.
After receiving the cells, if the cells are in poor condition or encounter problems during culture, please contact us as soon as possible on the same day for processing. Timely feedback on the cell status and cell photos on the same day is an important reference for after-sales tracking and processing, so please pay attention to it.
When the cells are just received, fetal bovine serum can be cultured with % for two or three days, which will help the cells recover as quickly as possible. After the cell status is stable, the % can be adjusted back.
(Among them, the strips are for adherent cells. For details on suspended cells, please refer to the cell operating instructions) h:r
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[Product packaging] h:r
Resuscitation culture Bottle (one bottle) or cryopreservation tube (two)
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[Product specifications] h:r
*
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[Security level] h:r
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[Product type] h:r
People
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[Organization source] h:r < br/>B cells
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[Cell morphology] h:r
Lymphoblastoid
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[Cell characteristics] h:r < br/>Suspension
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Selection and use of cell culture medium
M has simple ingredients and is the first choice for adherent cells.
DM is divided into high sugar type and low sugar type.
DM is suitable for cells with low cell density and difficult growth. Such as screening of hybrid cells, screening and culture of transformed cells after D transfection.
R is one of the most widely used culture media. The first choice for suspension cell culture, mainly for lymphocytes, but also suitable for other cells.
, culture medium: diffuse large B cells
Complete ingredients, good culture effect.
Medium: Suitable for mouse and human diploid cell culture.
Medium: Suitable for single cell isolation and culture and serum-free culture.
Mrq medium: Especially suitable for the growth of primary cells and cells that are difficult to culture. h:r
% ()
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【Cell Test】h:r
Cells do not contain H, HB, H, mycoplasma, bacteria, yeast and fungi
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[Cultivation conditions] h:r
See cell instructions for details
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[Passaging method] h:r
Recommendation: two days Change the medium once
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[Freezing conditions] h:r
See the cell instructions for details
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[Main literature] h:r
Please refer to relevant published articles for details
Trypsin digestion method for cell subculture
Subculture: refers to the transfer of cells from one culture bottle to another culture bottle at a ratio of: or: or above. nourish. Cell passage can be carried out in different ways according to different cells. The adherent cells were passaged by digestion method, and the partially adherent cells were passaged by direct pipetting or scraping with silicone soft scraper. Suspension cells can be passaged by adding an equal amount of fresh medium and then pipetting and dispersing directly, or by adding new medium by natural sedimentation method and then pipetting and dispersing for passage.
Experimental steps: ① Wash your hands with soap before entering the sterile room, and then wipe and disinfect your hands with % alcohol
② Observe the cell morphology under an inverted microscope to determine whether the cells are passaged and the number of times the cells need to be diluted. Preheat the culture medium at ℃.
③ The surface of the ultra-clean workbench should be clean and tidy, wipe it clean with % alcohol cotton balls
④ Turn on the ultraviolet lamp of the ultra-clean workbench to illuminate the surface for about m, turn off the ultraviolet lamp, and turn on the fan to clean the air and remove ozone
⑤ Light the alcohol lamp and take out the sterile test tube. Place the rubber tips on the Pasteur straw and the graduated straw and pass them through the flame of the alcohol lamp to slightly burn them and then insert them into the sterile test tube.
⑥ Disinfect the mouth of the culture solution bottle with % alcohol, pass it through the flame of the alcohol lamp and place it diagonally on a shelf next to the alcohol lamp.
⑦ Discard the old culture medium used to culture cells. If appropriate, use H solution to wash away the remaining culture medium, or rinse with a small amount of trypsin.
⑧ Add m trypsin to each large culture bottle and reduce the amount for small culture bottles. Cap the bottle and observe under an inverted microscope. When the cells retract the protrusions and become rounded, turn the culture bottle over immediately to release the cells from the trypsin, and then pour out the trypsin. Be careful not to let the cells fall off into the digestive juices early.
⑨ Add a small amount of fresh culture medium containing serum, repeatedly pipette the digested cells to detach and disperse them, and then add a certain amount of fresh culture medium containing serum (m/large bottle m) according to the number of distribution bottles. / vial) to prepare a cell suspension, diffuse large B cells
and then aliquot into new culture bottles. Cap the bottle, tighten it moderately and turn it slightly to facilitate access, and return the culture bottle to the incubator.
⑩ For suspension culture cells, this step is not performed. Centrifuge the cell suspension directly to remove the old culture medium supernatant, add fresh culture medium, and then dispense it into each bottle.
Precautions
① Pay attention to aseptic operation and prevent cross-contamination between cells when subculturing. All operations should be performed as close to the flame of the alcohol lamp as possible. It is best to work on only one type of cell at a time. Use one set of equipment for each cell type.
② Observe the cell morphology every day and grasp the standard of whether the cells are healthy: healthy cells have a plump shape, good refractive index, and can be passaged when they grow densely.
③ If signs of contamination are found in the cells, measures should be taken immediately. Generally, the contaminated cells should be discarded. If they must be rescued, antibiotic-containing or culture media can be added and washed repeatedly, and then a larger amount of antibiotics can be added to the culture medium, and Change culture medium frequently.
Establishment and maintenance of passaged cells: The maintenance of cell lines is achieved by changing the medium, changing the medium, re-passaging and cryopreservation of cell seeds.
Each cell line has its own characteristics. To maintain the line well, it is necessary to record the cell files, change the medium and pass the cells and manage the cell line well. Rr
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, diffuse large B cells
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H cells h:r
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OCI-LY10 diffuse large B cells