MSC-P5 Cells

The MSCP5 cell line, derived from murine skin keratinocytes, represents a vital tool for research in dermatology and cellular biology. This line is characterized by its robust expression of prostaglandin-H synthase 2 (PGHS-2), also known as cyclooxygenase-2 (COX-2), an enzyme critical in the prostaglandin biosynthesis pathway, which plays a pivotal role in inflammation and wound healing processes. Notably, MSCP5 cells exhibit a pronounced induction of PGHS-2 expression upon stimulation with phorbol 12-myristate 13-acetate (PMA), mimicking the cellular response to inflammatory conditions and hyperproliferative states of the epidermis.
This cell line offers a unique model for investigating the regulation of COX-2 expression and its implications in skin pathophysiology, including inflammation and carcinogenesis. The PMA-induced upregulation of PGHS-2 in MSCP5 cells provides a valuable system for studying the molecular mechanisms of keratinocyte response to inflammatory stimuli, the role of prostaglandins in skin diseases, and the potential therapeutic targeting of COX-2 in dermatological conditions.

Adherent

C3H

EMEM, w: 2 mM L-Glutamine, w: 1.5 g/L NaHCO3, w: EBSS, w: 1 mM Sodium pyruvate, w: NEAA (Cytion article number 820100c)

Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium.

After thawing, plate the cells at 5 x 10^4 cells/cm^2 and allow the cells to recover from the freezing process and to adhere 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.

A ratio of 1:4 to 1:8 is recommended

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.