Chicken embryo fibroblasts|100311 |BNCC

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UMNSAH/DF-1

Culture medium	BNCC338068
Description	DMEM-H Complete Medium (with 10% FBS)
Composition	90%DMEM-H+10%FBS
Growth conditions	culture temperature 39 ℃; Gas environment with 5% CO₂ and 95% air;
Subculture procedure	Resuscitation steps: ① Remove the cryotube from liquid nitrogen or -80 ℃ refrigerator and place it in PE gloves. Quickly immerse it in a 37 ℃ water bath, shake the cryotube to accelerate dissolution, and dissolve it completely within 1 minute; ② Add the dissolved cell solution into a centrifuge tube containing 9mL of complete culture medium on a super clean bench, centrifuge at 1000-1200rpm for 5 minutes, discard the supernatant, and resuspend the cells in 1-2mL of complete culture medium. ③ Add the cell suspension into a T25 bottle containing 5-6mL of complete culture medium and incubate in a culture incubator. Cell passage: ① Remove the old culture medium, wash twice with PBS, and add 1-2mL Trypsin (0.25% Trypsin + 0.02% EDTA); ② Under the microscope, observe the digestion process. When the cell edge shrinks and the adhesion becomes loose (you can use a straw to suck up some trypsin and gently blow it onto a certain part of the cell layer. The cell layer can be seen to have fallen off with the naked eye, indicating that digestion is complete. Otherwise, continue digestion). Directly suck out the trypsin, add 5-6mL of complete culture medium, gently blow the cell layer, blow it off, and disperse. ③ Divide the cell suspension into new T25 bottles at a ratio of 1:2, add appropriate complete culture medium, mix the cell suspension evenly, and culture it in the incubator.; ④ Pay attention to changes in the pH value of the culture medium and cell density, change the medium regularly (2-3 times a week), and repeat the passage operation or cryopreservation when the cell density reaches 80% -90%.
morphology	Epithelial-like，Polygonal，irregular edge，Monolayer adherent growth
Sharing mode	Public welfare sharing