Human embryonic kidney cells (ampho transfected)|BNCC353297 |BNCC

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Human embryonic kidney cells (ampho transfected)

Culture medium	DMEM-H complete medium: 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 solution into a centrifuge tube containing 9ml of complete culture medium in an ultra-clean table, centrifuge at 1000-1200rpm for 5min, 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, blow the cells with PBS, collect the cells, centrifuge at 1000rpm for 5 minutes, discard the supernatant, add 1-2mL of liquid media and blow well, divide the cell suspension into a fresh T25 bottle at a ratio of 1:2, add appropriate complete medium, mix the cell suspension evenly, and culture in an incubator. 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%. Notes: ① Cells are density-dependent; initiate the subculture (density reaches 80%), it is recommended to keep at the ratio 1:2 (mataining cell density) and preferentially in T25 flask. (2) If cultivated in sealed bottle, put it into an incubator after cap loose of for the culture bottle. (3) Cells tend to cluster. blow the cell suspension evenly during the recovery or subculture.
Growth conditions	Culture temperature 37 ℃; Atmosphere 5% CO2 + 95% air;
Growth characteristics	Adherent growth
Storage conditions	Liquid nitrogen
Type	Adherent growth
Safety level	1
morphology	epithelioid cells, diverse
application	3D cell culture High throughput screening Toxicology Vaccine research and development
Sharing mode	Public welfare sharing

Phoenix-AMPHO human embryonic kidney cells (ampo transfected)

Adherent, epithelioid cells

No. : 353297

Culture :37 ℃,5% CO₂ CM1-1 culture solution

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

Biosafety level : 1 , handle in ultra-clean 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 lose of cell viability.

Growth conditions :37 ℃,5% CO₂,CM1-1 culture solution. CM1-1 culture solution: 90% DMEM-H + 10% FBS. DMEM-H:DMEM high sugar culture solution, containing glutamine and sodium pyruvate.

Recovery steps:
(1)Prepare a new 100mm culture dish containing 12ml of the above culture medium;
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.
(2)Draw the solution with a sterile pipette, drip into a new culture dish, and mix it evenly by shaking clockwise;
(3)Put it into incubator (37℃，5%CO2）, change the media overnight, and it will grow up in 2-4 days.

Subculture / cryopreservation: remove old medium, and rinse twice with PBS, add 6ml of Trypsin, do not shake the culture dish during the period. Observed under the microscope, remove most of the trypsin(with 0.5ml left) when the cells detach. transfer it to the incubator for digestion about 2minute, and take it out.

(1)Subculture: terminate digestion with 12ml of CM1-1 culture media, aspirate and dispense into 3~6 culture dishes.

(2) Cryopreservation: terminate digestion with 6ml of cryopreservation solution （90%FBS+10%DMSO), aspirate and dispense into 6 frozen vials, and cryopreserve at -80 ℃ with 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 12hours, the cell adherence rate ≥ 80.0% in 70hours	adherence is observed in 18 hours, the cell adherence rate ≥ 80.0% in 66hours
cell morphology	adherent, epithelioid cells	in the CM1-1 culture solution, adherent, epithelioid cells, diverse and round, grow in blocks
attached figure
conclusion	good viability, and no abnormal cell morphology, qualified