Bovine endometrial epithelial cells|BNCC359233 |BNCC

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Bovine endometrial epithelial cells

Culture medium	DMEM-H complete medium (including 10% FBS):90% DMEM-H + 10% FBS
Subculture procedure	Recovery steps: ① Take out the frozen vial from liquid nitrogen or -80 ℃ and put it into PE gloves, quickly submerse into a 37 ℃ water bath pot, shake the frozen vial to accelerate dissolution, and it is advisable to dissolve all within 1 minute; (2) Add the dissolved cell liquid into a centrifuge tube containing 9ml of complete culture medium in an ultra-clean table, centrifuge at 1000-1200rpm/min for 5 minutes, discard the supernatant, resuspend the cells with 1-2ml of complete media. ③The cell suspension was then added to the T25 flask containing 5-6mL of complete medium and placed in an incubator for culture. Cell subculturing: ① remove the medium, rinse twice with PBS, add 1-2mL pancreatin (0.25%Trypsin +0.02%EDTA); (2) Observe the digestion under the microscope. When the cell edge shrinks and the adherent is loose (a pasteur pipette can be used to suck up some pancreatin blow somewhere in the cell layer, the cell layer can be seen to detach with naked eyes, I .e. digestion is completed, otherwise digestion is continued), and directly suck out pancreatin, add 5-6ml of complete medium, gently blow the cell layer, blow the cell layer off and blow it off. ③ Dispense the cell suspension into a fresh T25 flask as the ratio of 1:2, add appropriate complete culture medium, mix the cell suspension evenly, and culture in an incubator. (4) Pay attention to the change of pH of media and cell density, renew the medium regularly (2-3 times a week), and repeat the subculture or cryopreservation when the cell density reaches 80%-90%.
Growth conditions	Culture temperature 37 ℃; Atmosphere 5% CO2 + 95% air;
Growth characteristics	Adherent growth
Storage conditions	Liquid nitrogen
Safety level	2
morphology	Epithelial-like, polygonal, irregular margin, monolayer adherent growth, clean background, no vacuoles
Sharing mode	Public welfare sharing

Bovine endometrial epithelial cell BEND

Adherent, epithelial cell-like

No. 359233

Product format: 2ml frozen vial x 2, or T25 flask x 1

Biosafety level:1, handle in ultra-clear table or safety cabinet

Receiving notice: if any abnormality is found on the day of receipt, please contact the customer service within 24 hours. If it is overdue, it is deemed that the cells are well. For the frozen cells, it shall be stored in the refrigerator at -80 ℃ upon arrival. If they are not used for a long time, they shall be transferred to liquid nitrogen for storage overnight. Recovered cells in T25 culture flask, upon receipt, put the culture flask in the incubator for 4h, and then carry out handling procedure. During recovery, each vial shall be used up once and shall not be retained. After recovery, the cells can be passed on to the next generation and can be used normally. Please operate in strict accordance with this instruction, otherwise the replacement of cells are not be available in case of loss of cell viability.

Growth conditions: 37°C, 5% CO2, 90% DMEM-H+10% FBS. DMEM-H: DMEM high glucose medium containing glutamine and sodium pyruvate.

Recovery steps:

① Prepare a fresh 100mm culture dish containing 12ml of the above culture medium;

② Remove the frozen vial from liquid nitrogen or refrigerator at -80℃, thaw the vial in 37°C water bath for 1-2 minutes. Transfer the vial to biosafety cabinet for culture as soon as the contents are completely thawed.

③ Draw the solution with a sterile pipette, drip into a new culture dish, and mix it evenly by shaking clockwise;

④ Put it into incubator (37℃，5%CO₂）, change the media overnight, and it will grow up in 2-4 days.

Subculture/ cryopreservation: centrifuge the suspended cells at 110g for 3 minutes to collect the cells, rinse twice with PBS, add 2ml of 0.25%Trypsin+0.02%EDTA. Observe under the microscope, remove most of the trypsin(with 0.5ml left) when the cells detach. transfer it to the incubator for digestion about 1.5 minutes, and take it out.e: terminate digestion with 6ml of culture media, aspirate and then subculture at a ratio of 1:2;for cryopreservation, stop digestion with 3 mL of freezing solution (50% basal medium + 40% FBS + 10% DMSO), pipette evenly, divide into 3 cryopreservation tubes, and freeze at -80°C in a programmed cooling box.

Recovery record: According to the recovery instructions, the results of the cell recovery are reported as follows:

Item	quality standard	Recovery record
viability:	adherence is observed in 18 hours, the cell adherence rate ≥ 80.0% in 72 hours	adherence is observed in 18 hours, cell adherence rate ≥ 80.0% in 68 hours
cell morphology:	Adherence, epithelial cell-like	Adherent, epithelial-like, polygonal
attached:
conclusion: good viability, no abnormal cell morphology, qualified

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