PM-LGSOC-01 Cells

PM-LGSOC-01 is a cell line derived from peritoneal metastasis of a low-grade serous ovarian carcinoma (LGSOC). This cell line was established as part of a comprehensive research model that also included a patient-derived xenograft (PDX). The creation of PM-LGSOC-01 involved orthotopic engraftment via subperitoneal tumor slurry injection in SCID/Beige mice, leading to an early-stage transplantable peritoneal metastasis (PM)-PDX model. Histological analysis confirmed that both the PM-PDX and PM-LGSOC-01 cells retained the micropapillary and cribriform growth patterns typical of LGSOC, with tumor budding, and expression of markers such as PAX8 and WT1. Genetic analysis showed that the primary tumor, the PM, and the cell line share a KRAS c.35 G > T (p.Gly12Val) mutation, making this model relevant for studying LGSOC progression and treatment response, particularly in relation to the MAPK pathway.
PM-LGSOC-01 exhibits key characteristics relevant for preclinical research. It has a doubling time of approximately 42 hours in early passages, which decreased to 23 hours in later stages of cell culture, and has been maintained for over 100 in vitro passages. The cell line demonstrates epithelial morphology with cobblestone-like organization and high cell-cell adhesion. However, it shows limited response to platinum-based chemotherapy but is highly sensitive to paclitaxel (IC50: 6.3 ± 2.2 nM). Additionally, PM-LGSOC-01 is particularly sensitive to the MEK inhibitor trametinib (IC50: 7.2 ± 0.5 nM), both in vitro and in vivo, reflecting the impact of the KRAS mutation on therapeutic responses.
PM-LGSOC-01 serves as a valuable tool for investigating LGSOC, particularly in the context of drug resistance, tumorigenicity, and sensitivity to targeted therapies like MEK inhibitors. Its use in developing personalized treatment approaches for low-grade serous ovarian carcinoma is critical, given the poor responsiveness of LGSOC to conventional chemotherapy compared to high-grade serous ovarian carcinoma (HGSOC).

Adherent

60 years

Epithelial-like

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.

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 x 10^4 cells/cm^2

42 hours

Trypsin/EDTA and Ca2+/Mg2+ free phosphate buffer