Human plexiform neurofibroma cell Schwann cell|BNCC358003 |BNCC

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Human plexiform neurofibroma cell Schwann cell

Culture medium	hTERTNF1ipNF95.11bC Cell Special Medium: 89% DMEM + 10% FBS + 1% Sodium Pyruvate
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 5-6mL of complete medium and placed in an incubator for culture. Cell subculturing: ① remove the medium, rinse twice with PBS, and add 1-2mL pancreatin (0.25% Trypsin + 0.02% EDTA); (2) Observe the digestion situation under the microscope. When the cell edge shrinks and the adherent is loose (a pasteur pipette can be used to suck out a little pancreatin and gently blow somewhere in the cell layer, the cell layer can be seen to detach with naked eyes, that is, digestion is completed, otherwise digestion is continued), directly suck out pancreatin add 5-6ml of complete medium, gently blow the cell layer off. ③ Dispense the cell suspension into a fresh T25 flask as the ratio of 1:2, add appropriate complete culture 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	Adherent growth
Storage conditions	Liquid nitrogen
Type	Adherence
Safety level	1
morphology	Fibroblast/spindle
Separation substrate	Nerve: brachial plexus
application	Studying Schwann cell biology, diseases and tumors. These cells are valuable tools for dissecting neurofibromas and other possible disease-forming processes, as well as for the development of disease models and therapeutics.
Sharing mode	Public welfare sharing

human plexiform neurofibroma cells Schwann cells

adherent, Schwann cells

No. 358003

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 loss of cell viability.

Growth conditions:37 ℃,5% CO2,500 mLDMEM-H +56mLFBS + 5.6mL L-glutamine (200mM).

Recovery steps:

(1)prepare a new 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 3-5 days.

Subculture/Cryopreservation: suck out the old culture solution. after PBS cleaning twice, add 2mL(/100mm dish/T25 bottle) of pancreatin (0.25% Trypsin + 0.02% EDTA) and observe under a microscope. during this period, it is forbidden to shake the culture dish. when the cells just fall off, suck out most of the pancreatin, leave about 0.5mL, move to the incubator for digestion, and take out about 2min. Subculture is terminated by digestion with 6mL of culture solution, and cells are gently blown evenly, and then passaged at 1:3. For cryopreservation, 3mL of cryopreservation solution (90% culture solution + 10% DMSO) is used to terminate digestion, blow evenly, divide into 3 frozen storage tubes, and use a program cooling box to freeze storage at -80 ℃.

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 108hours	adherence is observed in 18 hours, the cell adherence rate ≥ 80.0% in 96hours
cell morphology:	Adherent, Schwann cell	Adherent, Schwann cell, spindle
attached：
Conclusion:	good viability, and no abnormal cell morphology, qualified

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