Human mammary epithelial cells|BNCC337734 |BNCC

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Human mammary epithelial cells

Culture medium	Special medium for mammary epithelial cells: MEGM serum-free medium
Subculture procedure	Recovery steps: ① One fresh 100mm plate containing 12mL of the above culture solution; (2) The frozen vial is taken out of liquid nitrogen or -80 ℃, bathed in water at 37 ℃ for 1~2min, and moved into the open it cabinet for recovery as soon as possible after complete dissolution; (3) using a sterile pipette to suck the dissolved solution into a fresh plate and shake well clockwise; (4) Put it into an incubator, change the medium. passage/cryopreservation: suck out the old culture solution, rinse twice with PBS, add 2mL(/100mm dish/T25 flask) pancreatin (0.25% Trypsin + 0.02% EDTA), observe under a microscope, do not shake the culture dish during this period, when the cells detach, suck out most of the pancreatin with about 0.5ml remained, move to the incubator for digestion, and take out in about 2min. Subculture is terminated by 6mL of 10%-containing FBS culture solution, and subculture at 1:2 after centrifugation at 1000rpm for 3min. For cryopreservation, resuspend the cells with 3mL of serum-free cell cryopreservation solution, mix evenly, dispense into 3 cryopreservation tubes, and freeze at -80 ℃ with a program cooling box.
Growth conditions	37 ℃;5% CO2 + 95% air;
Growth characteristics	Adherent growth
Storage conditions	Liquid nitrogen
Safety level	1
morphology	Epithelial cell-like, spindle-shaped, irregular margin, monolayer adherent growth, clean background
Sharing mode	Public welfare sharing

MCF-10A human mammary epithelial cells

Adherent, epithelioid cells

No.: 337734

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

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

Culture:37 ℃,5% CO₂ CM30-1 culture solution

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,CM30-1 culture solution. CM30-1 culture solution: 500ml MEpiCM +5ml MEpiCGS.

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 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 complete media supplemented with10%FBS, add 3ml of CM30-1 media after centrifuge, aspirate the cells, and dispense into 3-6 culture vessel.

(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
viability	adherence is observed in 18 hours, the cell adherence rate ≥ 80.0% in 107hours	adherence is observed in 12 hours, the cell adherence rate ≥ 80.0% in 100hours
cell morphology	adherent, epithelioid cells	in CM30-1 culture solution, adherent, epithelioid cells, round, short spindle
attached figure
conclusion	good viability, and no abnormal cell morphology, qualified