B2-17|342333 |BNCC

Essential Information Related Products

B2-17

Culture medium	BNCC338068
Description	DMEM-H Complete Medium (with 10% FBS)
Composition	90%DMEM-H+10%FBS
Growth conditions	37°C; 5% CO₂ + 95% air;
Attention	If any abnormality is found on the day of receipt, please contact customer service within 24 hours. Failure to do so will be considered as confirmation of good condition upon receipt.For cryovial format: Upon receipt, immediately store at -80°C. For long-term storage, transfer to liquid nitrogen overnight. During resuscitation, use the entire contents of one vial at a time; do not reserve any portion.For T25 flask format: Upon receipt, let the flask stand in the incubator for 2–3 hours before proceeding with routine cell handling.Please strictly follow the instructions above. Failure to do so may result in cell inactivation, and no replacement will be provided.
Subculture procedure	Thawing procedure: ① Remove the cryovial from liquid nitrogen or a -80°C freezer and place it inside a pair of PE gloves; quickly immerse it in a 37°C water bath, shaking the cryovial to accelerate thawing; it is best to ensure complete thawing within 1 minute; ② In a laminar flow hood, add the thawed cell suspension to a centrifuge tube containing 9 mL of complete culture medium; centrifuge at 1000–1200 rpm for 5 minutes; discard the supernatant; resuspend the cells in 1–2 mL of complete culture medium. ③ Add the cell suspension to a T25 flask containing 5–6 mL of complete medium, then place it in the incubator for culture. Cell Passaging: ① Aspirate the old culture medium, wash twice with PBS, then add 1–2 mL of trypsin (0.25% trypsin + 0.02% EDTA); ② Observe the digestion process under a microscope; when the cell edges begin to contract and the cells become loose from the surface, (you may use a pipette to draw up a small amount of trypsin and gently pipette it onto a spot on the cell monolayer; if the cell monolayer detaches visibly, digestion is complete; otherwise, continue digestion), directly aspirate the trypsin, add 5–6 mL of complete culture medium, and gently pipette the cell monolayer to dislodge and disperse the cells. ③ Transfer the cell suspension to a new T25 flask at a 1:2 ratio, add an appropriate amount of complete medium, mix the cell suspension thoroughly, and incubate in the incubator.; ④ Monitor changes in medium pH and cell density, and perform regular medium changes (2–3 times per week). When cell density reaches 80%–90%, repeat the passage procedure or cryopreserve the cells.
morphology	Epithelioid, polygonal, with irregular margins; monolayer adherent growth
Sharing mode	Public welfare sharing