Human multiple myeloma cells|BNCC338295 |BNCC

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Human multiple myeloma cells

Culture medium	IMDM complete medium: 80% IMDM + 20% FBS
Subculture procedure	Recovery steps: ① The frozen vial is taken out of liquid nitrogen or -80 ℃ refrigerator and put into PE gloves, quickly submersed into a 37 ℃ water bath pan, shaken the frozen vial to accelerate dissolution, and it is advisable to dissolve all within 1min; (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 medianatant. ③The cell suspension was then added to the T25 flask containing 6-8mL of complete medium and placed in an incubator for culture. cell subculture: ① transfer the cell suspension solution into a centrifuge tube, centrifuge at 1000rpm for 5min, and collect cells; (2) After double gentle rinsieT25 flaskt with PBS, add 1-2mL pancreatin (0.25% Trypsin + 0.02% EDTA); Observe the digestion situation under the microscope. When the cell edge shrinks and the adherent is loose (a pipette can be used to suck up some pancreatin, and gently blow somewhere in the cell layer, , 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 together and then dispensed into fresh T25 flask according to a ratio of 1:2, appropriate complete culture medium is added, the cell suspension is evenly mixed and cultured in an incubator. ④ Pay attention to the change of pH value of medium and cell density, change 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	B lymphocyte
Safety level	1
morphology	Lymphoblastoid, rounded, irregular margin, single cell, clean background, no pigment, no vacuoles
Separation substrate	Peripheral blood
application	There is no evidence that the cell can produce heavy chains (cytoplasmic or secretory).
Sharing mode	Public welfare sharing

RPMI 8226 human multiple myeloma cells

Suspension, lymphoblast-like

No. : 338295

Culture :37 ℃,5% CO₂CM2-1 culture solution

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

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 loss of cell viability.

Growth 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 8-10 days.

Subculture: the suspended cells were gently beaten and evenly distributed into 3-6 fresh culture medium dishes. After being shaken well, the cells were put into the incubator for culture

Cryopreservation: The suspended cells were transferred to the centrifuge tube for (110g, 3min) centrifugation，after centrifugationterminate 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	suspension growth rate ≥ 80.0% in 10days	inoculation with 20% cell solution, suspension growth rate reaches 40.0% in 18hours, and 80.0% in 8days
cell morphology	suspension, lymphoblast-like	CM2-1 culture medium, suspended, lymphoblast-like, round, few clusters grow
attached figure
conclusion	good viability, and no abnormal cell morphology, qualified