Mouse breast cancer cells|BNCC273810 |BNCC

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Mouse breast cancer cells

Culture medium	RPMI-1640 complete medium: 90% RPMI-1640 + 10% FBS
Subculture procedure	remove the medium, rinse twice with PBS, add 2mL(/100mm dish/T25 bottle) pancreatin (0.25% Trypsin + 0.02% EDTA), observe under a microscope, do not shake the culture dish during this period, when the cells just detach, absorb most of pancreatin, leave about 0.5mL, move to the incubator for digestion, and take out about 4min. Passage with 6mL of culture solution to stop digestion, gently blow uniform cells, after 1:2 passage;
Growth conditions	37 ℃,5% CO2,90% RPMI-1640 + 10% FBS. RPMI-1640:1640 culture solution, containing glutamine.
Storage conditions	For cryopreservation, the digestion is terminated with 3mL of cryopreservation solution (50% basic medium + 40% FBS + 10% DMSO), blown evenly, divided into 3 cryopreservation tubes, frozen at -80 ℃ with a program cooling box, and transferred to liquid nitrogen overnight for storage.
Type	Adherent growth
Safety level	1
morphology	adherent, epithelioid cells, polygonal, irregular
Separation substrate	4T1 is a 6-sulfur bird resistant cell line screened from 410.4 tumor strains without mutagenesis. When injected into BALB/c mice, 4T1 spontaneously produced highly metastatic tumors that could metastasize to the lungs, liver, lymph nodes, and brain, while developing an initial focus at the injection site. It is not necessary to remove the initial focus when inducing metastasis.
application	The tumor growth and metastatic spread of 4t1 cells from balb/c mice is very close to that of human breast cancer. This tumor is an animal model of advanced breast cancer in humans. The 4t1-induced tumor can be used as a postoperative model and a non-surgical model, because the 4t1-induced tumor has similar kinetic spontaneous metastasis in both models.
Sharing mode	Public welfare sharing

4T1 mouse breast cancer cells

Adherent, epithelioid cells

No. 273810

Culture:37 ℃,5% CO₂ CM2-1 culture solution

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.

Growthe conditions:37 ℃,5% CO₂,CM2-1 culture solution. CM2-1 culture solution: 90% RPMI-1640 + 10% FBS. RPMI-1640:1640 culture solution, containing glutamine.

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 5-7 days.

Subculture / cryopreservation: remove old medium, and rinse twice with PBS, add 2ml of 0.25%Trypsin+0.02%EDTA, 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 4minute, and take it out.

(1)Subculture: terminate digestion with 6ml of CM2-1 culture media, aspirate and dispense into 3~6 culture dishes.

(2) Cryopreservation: terminate digestion with 3ml of cryopreservation solution （90%FBS+10%DMSO), aspirate and dispense into 3 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
resurrection	adherence is observed in 18 hours, the cell adherence rate ≥ 80.0% in 168 hours	adherence is observed in 18 hours, cell adherence rate ≥ 80.0% in 144 hours
cell morphology	adherent, epithelioid cells	CM2-1 culture medium, adherent, epithelioid cells, polygonal, irregular
attached
Conclusion	good resurrection, no abnormal cell morphology, qualified