Human embryonic lung fibroblasts|BNCC340079 |BNCC

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Human embryonic lung fibroblasts

Culture medium	Special medium for IMR-90: 88% EMEM + 10% FBS + 1% NEAA + 1% L-glutamine.
Subculture procedure	remove the medium, rinse twice with PBS, add 2mL(/100mm dish/T25 flask) pancreatin (0.25% Trypsin + 0.02% EDTA), observe under a microscope, during which the culture dish is forbidden to be shaken. When the cells just detach, absorb most of pancreatin, leave about 0.5mL, move to the incubator for digestion, and take out in about 1min. Subculture with 6mL CM1-1 culture solution to stop digestion, gently blow uniform cells, after 3-6 dishes culture;
Growth conditions	37 ℃;5% CO2 + 95% air;
Growth characteristics	Adherent growth
Storage conditions	Liquid nitrogen
Safety level	1
morphology	Fibroblast-like, long fusiform, irregular margin, monolayer adherent growth, clean background, no vacuoles
Separation substrate	W.W. Nichols and colleagues established a human diploid fibroblast cell line IMR-90 from the lungs of a 16-week-old baby girl.
application	Division potential, viral susceptibility and other properties have been fully studied, so this cell can be used as a replacement for WI-38 or other standard human lung cell lines. It has been reported that this cell can double 58 times before showing senescence.
Sharing mode	Public welfare sharing

IMR-90 human embryonic lung fibroblasts

Adherent, fibroblast-like cells

No. : 340079

Growth :37 ℃,5% CO₂ CM1-1 culture solution

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 lose of cell viability.

Growth conditions :37 ℃,5% CO₂,CM1-1 culture solution. CM1-1 culture solution: 90% DMEM-H + 10% FBS. DMEM-H:DMEM high sugar culture solution, containing glutamine and sodium pyruvate.

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/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 1minute, and take it out.

(1)Subculture: terminate digestion with 6ml of CM1-1 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
viability	adherence is observed in 18 hours, the cell adherence rate ≥ 80.0% in 256hours	adherence is observed in 18 hours, the cell adherence rate ≥ 80.0% in 216hours
cell morphology	adherent, fibroblast-like cells	in CM1-1 culture solution, adherent, fibroblast-like cells, slender
attached figure
conclusion	good viability, and no abnormal cell morphology, qualified