SW948 human colon carcinoma cells
,SW-948
Item number: YJ-h356 (STR identifier )
Price: 1800.0
Specification: 1*106
Cell introduction
Human colon adenocarcinoma cell SW948 [SW-948], positive for keratin immunoperoxidase staining. The cells were positive for the expression of c-myc, K-ras, H-ras, N-ras, myb and fos oncogenes and the expression of N-myc and sis oncogenes was not detected.
STR identification result For: Amelogenin: X; CSF1PO: 12; D13S317: 10.11; D16S539: 11.12; D5S818: 11; D7S820: 9.11; THO1: 6.9.3; TPOX: 8.11; vWA: 16.18< /p>
Cell Functions
1) Source: white female 81 colon
2 ) Morphology: epithelial-like adherent growth
3) Content: >1x10^6 cells
4) Specification: T25 bottle or 1mL cryovial wrapper
5 ) Purpose: For research purposes only.
Shipping and storage
Dry ice transport and resuscitation of viable cells
(1) 1ml cryopreservation tube packed with dry ice for transportation, upon receipt, keep in -80 degrees refrigerator overnight, then transfer to liquid nitrogen or resuscitate immediately, if the dry ice appears to have evaporated. If the vial cap falls off, is damaged or the cells are contaminated, please contact us immediately.
(2) The surviving cells resuscitated in T25 flasks are shipped at room temperature. Upon receipt, please follow the treatment method of cells upon receipt.
Process after receiving cells
1) close After reaching the cells, disinfect the flask wall with 75% alcohol and place the T25 flask in an incubator for approximately 2-3 hours of 37°C damaged, there is liquid overflow and the cells are contaminated, please take a photo and contact us in time.
2) Please put in 4 or 5X Confirm the condition of the cells under a microscope and maAt the same time take 2-3 photos of the newly received cells (10×, 20×) and keep a photo of the appearance of the culture flask as a basis for the state of the cells upon receipt after sale.
3) Adhering Cells: The cells are incubated for 2-3 placed in an incubator at 37°C for hours and the growth and adhesion of the cells observed under a microscope. Some adherent cells fall off due to vibration during express delivery and form agglomerates after falling off. If the growth density of the cells observed under the microscope is less than 60%, the perfusion medium in the culture flask can be removed (if there are any loose cells, they should be recovered by centrifugation, resuspended and poured into the original culture flask). and the newly prepared 6-8 ml of complete medium was placed in the cell culture incubator for further kweken. When the cell growth density reaches 70%-80% or more, the cells can be subcultured. If the cells fall off during the passaging process due to transport and vibration, they should be recovered by centrifugation.
4Notes: The medium used for transport (perfusion medium) can no longer be used to culture cells. Replace with a new complete medium prepared according to the cell culture conditions in the cell culture manual. It is recommended that T25 culture flasks be passed 1:2 for the first pass after receiving the cells.
One. Preparation of medium and culture conditions
1) Prepare L15 (recommendedYJ-0009) medium; high quality fetal bovine serum, 10 %; double antibody, 1%.
<p style=\"margin: 40px 0px 10px; padding: 0px; line-height: 28px;\" 2) Culture conditions: Gas phase: air, 100%;The cell culture cannot be fed with CO2. If there is no condition to use an incubator with 100% air to prepare gas phase, airtight sealable lid can be used T25 culture flasks were used for the culture During the culture process, the cells were removed from the incubator and aired for 1-2 times Temperature: 37 degrees Celsius, the humidity of the incubator is 70%-80% Reference subculture cycle: 5-7 days, average change every 2-3 days.
3) Freezing solution: 90% serum, 10% DMSO, ready to use.
two. Cell handling:
1) Restore frozen cells:
Shake the cryopreservation tube containing 1 ml of cell suspension in a 37°C water bath to rapidly thaw, add to the centrifuge tube containing 4-6 ml of complete medium and mix well. Centrifuge at 1000 RPM for 3-5 minutes, discard the supernatant and resuspend the cells in complete medium. Then add the cell suspension to a culture flask (or dish) containing 6-8 ml of complete medium and culture overnight at 37°C. The next day, cell growth and cell density were observed under a microscope.
2) cells Subculture: When the cell density reaches 80%-90%, subculture can be executed.
< strong> For stickersThe passage of parietal cells can refer to the following methods:
1. Discard the culture supernatant and wash the cells 1-2 times with PBS without calcium and magnesium ions.
2. Join 0.25% (w/v) trypsin- 0.53 mM EDTA in culture flasks (1-2 ml for T25 flasks, 2-3 ml for T75 flasks), and placed in a 37°C incubator for 1-2 minutes (Indigestible cells can be appropriately extended digestion time) and then observe the digestion of the cell under a microscope. If most of the cells become round and fall off, quickly return them to the operating table, tap the culture flask a few times, then add 3 -4 ml of medium containing 10% FBS to stop digestion.
3. Lightly beat and aspirate, centrifuge at 1000 RPM for 3-5 minutes, discard the supernatant, add 1-2 ml of culture medium and blow evenly. Distribute the cell suspension in new T25 flasks at a ratio of 1:2, add 6-8 ml of new complete medium configured according to the instructions to maintain cell growth and vitality, and follow up passage according to the actual situation at 1: 2~1:5 ratio.
3) cells Cryopreservation: After receiving the cells, it is recommended to freeze a batch of cell seedsduring the first 3 culture passages for later experiments.
The following T25 bottle is an example;
1. Collect the digested cells in a centrifuge tube according to the process of cell subculture when freezing the cells, and use a hemocytometer to count them to determine the freezing density of the cells. The recommended freezing density for general cells is 1 × 10^ 6~1 ×107 live cells/ml.
2.1000rpm Centrifuge for 3-5min, remove the supernatant. Resuspend the cells with the prepared cell cryopreservation medium and divide into 1ml cryopreservation medium with 1×10^6~1 ×107 viable cells/ml Divide the cells into a cryopreservation tube and mark the name, generation, date and other information.
Place the cells to be frozen in a programmed cooler, place in a -80°C refrigerator overnight, then transfer to a container of liquid nitrogen for storage.At the same time, note the position of the cryotube in the container of liquid nitrogen for later reference and use.
Precautions
1. Upon receipt of the cellIf the dry ice has evaporated, the caps of the cryotubes have fallen off, are damaged or the cells are contaminated, please contact us immediately.
2. Do not open the bottle cap until the cells are reached, wipe the bottle body with alcohol and place it in the incubator for 2-4 hours (depending on the cell density) to stabilize the cell state. Then observe the growth of the cells under an inverted microscope and take pictures of the cells at different magnifications (it is recommended to take a picture of the overall appearance when collecting the cells, observe the color of the medium and check for leakage, then take pictures of the condition of the cells under the microscope, 100*, 200* each), observe and note if the cells are contaminated during transport. as the basis for our sales.
3. Because The cell status is affected by many factors such as environment, operation and transportation, so the company only guarantees the cell status within one week after the customer receives the cell, so the customer must show the proof of the time of receipt of the cell and the customer\'s receipt time and after the problem is discovered, the communication time of the customer service personnel proves that the interval between periods cannot exceed 7 days.
4. All animal cells are considered potentially biohazardous and should be handled in a second level biosafety bank, paying attention to protection. All waste fluids and containers that have come into contact with the cells must be sterilized before they can be used.and are thrown away.
5. Customer If you have any technical questions during the cell culture process, you can contact the after-sales technical service, and we will answer them at any time.
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