AT-1 Cells

The AT-1 cell line is a subclone of the parental R3327 rat prostate adenocarcinoma cell line. This particular cell line was derived from the Dunning model, which is a well-established model used to study prostate cancer. The AT-1 subclone is characterized by its relatively slow growth rate and low metastatic potential compared to other subclones derived from the same tumor, such as the MatLyLu (high metastatic potential) and AT-2 (moderate metastatic potential) cell lines. This makes the AT-1 cell line particularly useful for studies focused on the biology of non-metastatic or minimally invasive tumors.

In research settings, the AT-1 cell line has been utilized extensively to investigate the mechanisms of prostate cancer progression and to assess the efficacy of therapeutic agents. The cells generally exhibit a cuboidal morphology and are adherent. They have been shown to respond to hormonal manipulations, which mimics the hormonal responses seen in clinical prostate cancer. Studies using the AT-1 cell line have contributed to a better understanding of the interactions between tumor cells and the microenvironment, angiogenesis, and the molecular pathways involved in cancer progression. Importantly, the AT-1 cell line has been a valuable tool in the development of therapeutic strategies that are less focused on metastasis and more on primary tumor growth and local invasion.

Adherent. The cells form clusters in soft agar and can be adapted to suspension growth

Epithelial-like

RPMI 1640, w: 2.1 mM stable Glutamine, w: 2.0 g/L NaHCO3 (Cytion article number 820700a)

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 4 x 10^4 cells/cm^2 and allow the cells to recover from the freezing process and to adhere for at least 48 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 1:3 to 1:6 is recommended