Human embryonic lung fibroblasts|BNCC340264 |BNCC

Essential Information Certificate Related Products

Human embryonic lung fibroblasts

Culture medium	DMEM-H/F12 complete medium: 90% DMEM-H/F12 + 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 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	fibroblast
Safety level	1
morphology	Fibroblast-like, long fusiform, short fusiform, neat edges, monolayer adherent growth, clean background, no pigment, no vacuoles
Separation substrate	Lung
Sharing mode	Public welfare sharing

1.Name:HEL299

2. No.:340264

3. growth properties : □ wall ;■ suspension ; □ semi-suspension and semi-wall

4. Growth conditions:

culture medium	90% RPMI-1640
serum	10% FBS(Diagnovum)
temperature	37 ℃
air condition	5% CO₂,95% AIR
growth algebra	P4-5
frozen storage conditions	culture medium 50%, serum 40%, DMSO 10%

5. composition:

composition	specifications
a bottle of cells	T25
cell culture and operating instructions	1 copy

Receiving notice:

1 Upon arrival, it is suggested to sit the cells in a incubator for about 4 hours, and then renew the media for recovery or subculture according to the cell density.
2 Collect the cells by centrifuge, re-suspend the collected cells with 10ml of complete media, transfer to fresh culture flask/vessel and cultivate overnight, and dispense into separate vials for subculture according to the cell density.

3 Clustered growth: agitate the culture flask to disperse the clustered cells and continue to recover or subculture.

Recovery and subculture procedure of cells ( under strict aseptic conditions )

1. The suspension cells is normally handled by changing half of the solution and sub-cultured in separate flasks, that is, transfer half of the suspension liquid to a fresh flask/vessel, add appropriate complete medium and culture it in the incubator; It can also be sub-cultured in separate flasks according to cell density.
2 Pay attention to the change of pH value of medium and cell density, renew the medium regularly, and repeat the subculture or cryopreservation when the cell density reaches 70%-80%.
Special attention: (if handle in public laboratory or first time to culture the cells, it is recommended to add penicillin streptomycin into the media)
1 Upon the receipt of the cells, renew the solution with fresh 10% FBS medium as soon as possible. It is not recommended to use the medium used for transportation in the original flask.
2 Please take photos and contact the technicians in time if the culture flask leaks upon the receipt of cells.
3 Any compliant on the cells, please take photos and contctat our technicians.

Download certificate