P-815 Cells

Mouse cell lines

Hematopoietic

Mast cell

Male

Round cells

DMEM, w: 4.5 g/L Glucose, w: 4 mM L-Glutamine, w: 1.5 g/L NaHCO3, w: 1.0 mM Sodium pyruvate (Cytion article number 820300a)

Start cultures at 2 x 10^5 cells/ml and maintain between 1x10^5 and 1x10^6 cells/ml. Subculture by transferring an appropriate aliquot of the suspension into new flasks filled with fresh cell culture medium. Detach adherent cells by washing with PBS first and incubation with Accutase at ambient temeratur 8-10 minutes. Combine with the nonadherent cells and distribute into new cell culture flasks.

After thawing, allow the cells to recover from the freezing process for at least 24 hours.

Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

Upon receipt, either store the cryovial immediately at temperatures below -150° C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required.

For immediate culturing, swiftly thaw the vial by immersing it in a 37° C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains.

Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently.

Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth.

Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

When planning to store a cryovial in liquid nitrogen for future thawing, it is mandatory to adhere to stringent safety measures. Appropriate protective gloves and clothing are essential, and the use of a face mask or safety goggles is required during the transfer of frozen samples to or from the liquid nitrogen tank. This is to mitigate the risk of injury from potential cryovial explosions upon removal, which can result in the projection of sharp fragments.

We stand by the promise of delivering products with high cell viability and robust culture performance. To achieve the best results, please make sure you follow the storage and culture instructions detailed in the product information sheet closely. Your adherence to these guidelines is key to success.

1 to 2 x 10^6 cells/ml

Deutsch:
Universitäre Kunden: Für den Erwerb ist ein Material Transfer Agreement oder eine Limited Use Label License auszufüllen.
Kommerzielle Kunden: Für den Erwerb ist ein Material Transfer Agreement oder ein Master Supply Agreement auszufüllen.
Nach eingegangener Bestellung werden ihnen alle relevanten Dokumente zugeschickt.

English:
University Customers: A Material Transfer Agreement or Limited Use Label License must be completed for purchase.
Commercial Customers: A Material Transfer Agreement or Master Supply Agreement must be completed for purchase.
After the order is received, all relevant documents will be sent to you.