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Shanghai Kanglang Biotechnology Co., Ltd
TE-6 human esophageal cancer cells
NegotiableUpdate on 05/06
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Overview
TE-6 human esophageal cancer cells are adherent cells with epithelial cell like morphology, and the tissue originates from the esophagus.
Product Details
1、 Basic cellular properties | |
cell name |
TE-6 human esophageal cancer cells |
Cell nickname |
TE-6 |
Product Item Number |
KL-C0007H |
source organism |
person |
Age and gender |
Male 71 years old |
source of organization |
esophagus |
Growth characteristics |
Wall attached growth |
cell morphology |
Epithelial like cells |
Background Introduction |
- |
Biosafety level |
- |
Cell specifications |
1 × 10 ^ 6 cells/T25 culture bottle or 1mL cryovial packaging |
Mycoplasma testing |
nothing |
culture medium |
RPMI1640+10 �S |
culture conditions |
Gas phase: 95% air+5% carbon dioxide; Temperature: 37 ℃ |
Freezing conditions |
Serum free cryopreservation solution, stored in liquid nitrogen |
Doubling time |
~24-30 hours |
Cell screening:
This cell has been stably transfected with LUC, and as the number of cell passages increases, its LUC fluorescence intensity gradually decreases. If the experiment requires maintaining fluorescence intensity, puromycin can be added for further screening.
After screening with 20 μ g/ml puromycin, cells can maintain 10 μ g/ml puromycin in normal culture
2、 Processing after cell reception
1) After receiving the cells, disinfect the bottle wall with 75% alcohol and place the T25 bottle at room temperature for about 1 hour. If you find that the culture bottle is damaged, there is liquid overflow, or the cells are contaminated, please take photos and contact us promptly.
2) Please confirm the cell status under a 4 or 5X microscope, and take 2-3 photos (10 ×, 20 ×) of the newly received cells, as well as one photo of the appearance of the culture bottle, for retention as a basis for the cell status upon receipt during after-sales service.
3) Adherent cells: Cells are left at room temperature for 1 hour, and their growth and adherence are observed under a microscope. Some adherent cells may detach and form clusters due to vibration during express delivery. If the growth density of cells observed under the microscope is below 60%, the culture medium in the culture bottle can be removed (if there are unattached cells, they need to be recovered by centrifugation and resuspended into the original culture bottle), and 6-8mL of newly prepared complete culture medium can be added and placed in the cell culture box for further cultivation. If the cell growth density reaches 70% -80% or more, the cells can be passaged. During the passage process, if cells shed due to transportation vibrations, they need to be recovered by centrifugation.
4) Method for handling cell detachment during transportation: Collect and centrifuge the detached cells, discard the supernatant, wash it with PBS, centrifuge and discard the supernatant, add 1ml of enzyme to the centrifuge tube for digestion for about 20 seconds, add complete culture medium to terminate digestion, centrifuge and flatten again, and the remaining adherent cells are passaged according to the normal adherent cell passaging method.
5) Note: The transport medium (infusion medium) cannot be used to culture cells anymore. Please use the newly prepared complete culture medium according to the instructions for cell culture conditions to culture cells. After receiving the cells, it is recommended to subculture them for the first time in a T25 culture bottle at a ratio of 1:2.
IIITE-6 human esophageal cancer cellsTraining operation
1) Resuscitation of cells: The following cell culture cryopreservation treatment is for reference only. The specific operating steps are mainly based on the product manual provided with the goods. Quickly shake and thaw the cryopreservation tube containing 1 mL of cell suspension in a 37 ℃ water bath, and mix 4 mL of culture medium evenly. Centrifuge at 1000 rpm for 3 minutes, discard the supernatant, add 1-2 mL of culture medium, and blow well. Then add all cell suspensions to a culture bottle containing an appropriate amount of culture medium and culture overnight (or add the cell suspensions to a 6 cm dish, add about 4 mL of complete culture medium, and culture overnight). On the third day, change the fluid and check the cell density.
2) Cell passage: If the cell density reaches 80% -90%, passage culture can be carried out.
a) Discard the culture supernatant and rinse the cells 1-2 times with PBS that does not contain calcium or magnesium ions.
b) Add 1 mL of digestion solution (0.25% Trypsin-0.02-TA) to the culture bottle, allowing the digestion solution to infiltrate all cells. Place the culture bottle in a 37 ℃ incubator for 1-3 minutes (depending on the digestion status of the cells), and then observe the digestion status of the cells under a microscope. If most of the cells become round and fall off, quickly take them back to the operating table, tap the culture bottle a few times, and add 2-3 mL of complete culture medium to terminate digestion.
c) Gently mix well and transfer to a sterile centrifuge tube. Centrifuge at 1000 rpm for 5 minutes, discard the supernatant, add 1-2 mL of culture medium, and blow well.
d) Divide the cell suspension into a new dish or bottle containing 8 mL of culture medium at a ratio of 1:2, and place it in a culture incubator for cultivation.
Attention points for passaging adherent cells:
Be sure to wash with PBS once.
Observe the cells several times to control the time. Do not stop digestion and blow them down before the cells have basically shed, as this makes it easier for the cells to differentiate and die.
Do not constantly blow and beat cells
3) Cell cryopreservation: When the cells are in good growth condition, cell cryopreservation can be performed. Taking T25 bottles as an example below;
a) Collect cells and cell culture medium, transfer them into sterile centrifuge tubes, centrifuge at 1000 rpm for 4 minutes, discard the supernatant, wash with PBS, discard PBS, and perform cell counting.
b) Add serum-free cell cryopreservation solution according to the number of cells, make the cell density 1 × 10 ^ 6~1 × 10 ^ 7/mL, gently mix well, and freeze 1mL of cell suspension in each cryopreservation tube, paying attention to labeling the tubes properly.
c) Place the cryovial in a -80 ℃ freezer and transfer it to liquid nitrogen for storage after 24 hours. Record the location of the freezer for future retrieval.
4、 Cultivation precautions
1) If there is a problem with the cell, it should be resent
a) Various problems encountered during cell transportation, such as cell loss, bottle damage, severe leakage of culture medium, etc., require resending;
b) Regarding the issue of cell contamination, please provide us with authentic experimental results within 3 days of receiving the product, verify them, and resend them;
c) After allowing the cells shipped at room temperature to stand for 2 hours, and after 2 days of recovery from dry ice freezing, the vast majority of the cells did not survive. (True and clear photos of the cell status should be provided), they should be resent;
d) After 2 days of revival of cells stored in dry ice for shipment or 2 hours of standing at room temperature for shipment without opening, if contamination occurs, resend;
e) Regarding cell viability issues, please provide us with real experimental results within 7 days of receiving the product, identify cell viability using the trypan blue staining method, and provide daily cell photos for verification before resending;
f) Please take photos on the day the cells are received, as well as on the 2nd and 3rd day. If you do not inform us within 3 days, it will be considered that the product is qualified. If there is a problem within 4-7 days, please provide photos taken 3 days before receiving the cells, photos of the cell problem, and detailed steps of the cell related operations, and communicate with the technical personnel. If the technical personnel determine that it is our responsibility, it will be resent.
2) The situation where cells have problems and are not resent
a) Customer operation causing cell contamination, not resend;
b) Serious operational errors by the customer resulting in poor cell status will not be resent;
c) If the cell state is not good due to our recommended cell culture system, it will not recur;
d) The cell status is not good, and no clear and authentic photos of the cell status from the first 3 days of culture have been provided. It will not be resent;
e) If other treatments during cell culture cause problems with the cells, they will not be reproduced;
f) If the time for receiving cells and discovering problems and communicating with customer service personnel is more than 7 days, it will not be resent;
g) It depends on the specific situation.
5、 Precautions
1. All animal cells are considered to have potential biological hazards and must be operated in a secondary biosafety platform. Please pay attention to protection, and all waste liquids and containers that have come into contact with these cells must be sterilized before disposal.
2. It is recommended to always use protective gloves, clothing, and a face mask when reviving frozen cells. Attention: The cryotube immersed in liquid nitrogen will leak and gradually fill with liquid nitrogen. When thawing, the conversion of liquid nitrogen into gas phase may cause the container to explode or the lid to be blown off with dangerous force, resulting in flying debris and causing personal injury.
3.This cell is for scientific research purposes only! The manual is for reference only. The actual delivery manual shall prevail!
