Olaparib-resistant PEO1-OR cell line

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Olaparib-resistant PEO1 derivative carrying a BRCA2 secondary mutation that restores full-length BRCA2, for studying acquired PARP inhibitor resistance in HGSOC

Contributors

Inventor Institute
Aneta Rogalska, Łukasz Biegała, Agnieszka Marczak University of Lodz
SKU: 162283
Product description: PEO1-OR is an olaparib-resistant derivative of PEO1 generated by continuous, stepwise exposure to increasing concentrations of olaparib. The model carries a secondary BRCA2 c.4964A>T mutation that reverses the truncating BRCA2 defect present in parental PEO1 and results in expression of full-length BRCA2. Whole-exome sequencing and functional characterisation also showed increased expression of homologous recombination repair factors, including BRCA1, BRCA2 and RAD51, consistent with restoration of homologous recombination capacity. These features make PEO1-OR a mechanistically characterised model of acquired PARP inhibitor resistance, particularly useful for investigating BRCA2 reversion-mediated resistance and evaluating approaches to overcome olaparib resistance.
Alternate name: PEO1-OR; PEO1-OR cell line; PEO1-OR cells; olaparib-resistant PEO1-OR ovarian cancer cell line
Gender: Female
CRISPR: No
Production details: PEO1-OR cell line is an olaparib-resistant subline of PEO1 cell line developed by addition of a stepwise increase in olaparib to the growth medium of the parental line.
Additional notes: PEO1-OR cells are grown and subcultured continually in the absence of olaparib. Please be aware that the originating laboratory of the PEO1 cell line acknowledges that PEO1 is both genetically unstable and derived from a heterogeneous population that was already present in the patient at the time of biopsy. This is evident in the literature (Cooke et al., 2010). Genetic differences within the PEO1, PEO4 and PEO6 cell lines suggest that PEO4 and PEO6 are not direct descendants of PEO1 but have diverged from a common ancestor.
Parental cell line: PEO1
Model description: Key mutations determined by whole-exome sequencing using NGS: TP53 (c.731G A; p.G244D); BRCA2 (c.[4964A>T; 4965C>G], p.Y1655L))
Disease: Cancer
Related tools: 151672 ; 151784 ; 151673 ; 151674
Cat. #: 162283
Provenance and Ownership: The Olaparib-resistant PEO1-OR cell line is owned by University of Lodz. CancerTools exclusively supplies the Olaparib-resistant PEO1-OR cell line under licence from University of Lodz and is the authorised source for its distribution. Sourcing Olaparib-resistant PEO1-OR cell line directly from the only licensed supplier ensures authenticity, provenance and compliance with all applicable licensing and research use requirements.
Cancer types: Ovarian cancer
Organism: Human
Tissue: Ovary
Gender: Female
Morphology: Epithelial
Growth properties: Adherent
Primary citation: Biegala et al. (2023) Cells, 12(7), 1038, PMID: 37048111
Initial handling information: Thaw cells rapidly in a 37°C water bath for up to 2 minutes, until a small amount of ice remains. Slowly add 1 mL of pre-warmed growth medium (37°C) to the thawed cells. Gently reconstitute the cell pellet by pipetting, then transfer the resuspended cells to a centrifuge tube and add 10 mL of pre-warmed growth medium (37°C). Pellet the cells by centrifugation (300×g, 5 min, RT) to remove the storage medium containing DMSO. After the centrifugation, cell pellet should be visible. Aspirate the supernatant, add 1 mL of growth medium, and gently resuspend the cells. Transfer the cells to into the appropriate culture vessel (e.g. T75 flask) containing medium that has been equilibrated in the incubator for a few minutes (37°C, 5% CO2). Optionally, take a small aliquot to assess cell viability. Rock the flask to distribute the cells evenly and place it in the incubator (37°C, 5% CO2). Thawed cells should be plated at high density to optimize recovery. Examine the cells microscopically after 24 hours and sub-culture as necessary.
Format: Frozen
Shipping conditions: Growth medium (RPMI 1640 culture medium + 2 mM glutamine + 25 mM HEPES + 10% heat inactivated FBS) + 10% sterile DMSO
Growth medium: RPMI 1640 culture medium with 2 mM glutamine and 25 mM HEPES supplemented with 10% heat inactivated FBS
Subculture routine: Split sub-confluent cultures (70–90%) every 2 to 4 days using a 0.25% trypsin-EDTA solution. Briefly, remove growth medium, wash cells with PBS without Ca2+/Mg2+, add trypsin-EDTA solution (approximately 1.5–2 mL per 75 cm2 of surface area), place flask in the incubator and leave it for 5–10 minutes (37°C, 5% CO2). Once the cells are detached, add fresh growth medium, and transfer the required number of cells to a new flask. Optionally, after trypsin inactivation, centrifuge cells (300×g, 5 min, RT), resuspend cell pellet in growth medium and passage required number of cells to a new flask. Recommended culture vessel: cell culture treated flasks with filter caps. As a general guide, 70–90% confluent cells should be split in approximately 1:4, 1:6 or 1:12 ratios to reach appropriate confluency after 2, 3 or 4 days. However, splitting ratio may vary depending on cell line handling and culture conditions. The complete formulation of the recommended trypsin-EDTA solution is: KCl (75 mg/L), KH2PO4 (60 mg/L), NaHCO3 (350 mg/L), NaCl (8000 mg/L), Na2HPO4 · 7H2O (90 mg/L), D-glucose (1000 mg/L), EDTA–4Na · 2H2O (380 mg/L), phenol red (10 mg/L), trypsin (2500 mg/L), pH 7.2 to 8.
Temperature: 5% CO2
Atmosphere: Dry ice
Storage medium: Liquid nitrogen, 2×10^6 cells/vial
Mycoplasma free: Yes
Biosafety level: 2
References:

Biegala et al. (2023) Cells, 12(7), 1038, PMID: 37048111

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