Human megakaryocytic leukemia cell|BNCC359444 |BNCC

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Human megakaryocytic leukemia cell

Culture medium	RPMI-1640 complete medium: 90% RPMI-1640 + 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 6-8mL of complete medium and placed in an incubator for culture. Cell subculturing: ① transfer the cell suspension solution into a centrifuge tube, centrifuge at 1000rpm for 5 minutes to collect cells; (2) rinse the T25 flask twice with PBS, add 1-2mL pancreatin (0.25% Trypsin + 0.02% EDTA); Observe the digestion situation under the microscope. When the edge of the cell shrinks and the adherent is loose (a pasteur pipette can be used to suck up some pancreatin , the cell layer can be seen to detach with the naked eye, that is, digestion is completed, otherwise digestion is continued), directly suck out pancreatin, add 5-6ml of complete culture medium, gently blow the cell layer off. (3) The cells obtained from the above two steps are evenly mixed and then dispensed into fresh T25 flasks as a ratio of 1:2. add appropriate complete medium, mix the cell suspension evenly, and culture in an incubator. ④ Pay attention to the change of pH of medium 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	semi-adherent and semi-suspension growth
Storage conditions	Liquid nitrogen
Type	Megakaryocytic leukemia cell
Safety level	1
morphology	Long fusiform, round, irregular margin, single cell, few clusters, clean background, no vacuoles
Separation substrate	megakaryocytic leukemia
Sharing mode	Public welfare sharing

Human megakaryocyte leukemia cell Dami

Semi-adherent growth, megakaryocyte

No. 359444

Product format: 2ml frozen vial x 2 or centrifuge tube 15mL ;

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. For centrifuge tube 15mL.,please place the original centrifuge tube into incubator fir 4 hours after receiving it and then handle routinely for the cells . 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 ℃,5% co ₂,90% RPMI-1640 + 10% FBS. RPMI-1640:1640 culture solution, containing glutamine.

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 4-6 days.

Subculture: Gently pipet the cultured suspension cells evenly, distribute them into 2~3 fresh culture medium dishes, shake gently and put them into the incubator for culture.

Cryopreservation: centrifuge the suspended cells at 110g for 3 minutes to collect the cells, After centrifugation, resuspend the cells with 3 mL of freezing solution (50% basal medium + 40% FBS + 10% DMSO), pipette evenly, divide them into 3 cryopreservation tubes, and freeze them at -80 °C in a programmed cooling box, overnight. Transfer to liquid nitrogen storage.

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

Item	quality standard	Recovery record
viability:	suspension growth rate ≥ 80.0% in 150 hours	inoculation with 30% cell solution, suspension growth rate reaches 40.0% in 18 hours, and 80.0% in 96hours
cell morphology:	Semi-adherent growth, megakaryocyte	Semi-adherent growth, megakaryocyte, round
attached：
conclusion: good viability, no abnormal cell morphology, qualified ;

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