LS513 Cells

Categories: Intestine cancer cell lines Description: The LS513 cell line is a well-characterized colorectal carcinoma model derived from a primary tumor biopsy taken in 1985 from a 63-year-old Caucasian male patient. The tumor was classified as a Dukes' C mucin-secreting cecal carcinoma located at the Bauhin valve. LS513 cells are adherent in nature and have demonstrated multidrug resistance (MDR), making them a valuable model for studying drug resistance mechanisms in colorectal cancer. These cells exhibit a 30% colony-forming efficiency in methylcellulose and are tumorigenic in nude mice, further validating their use in oncogenic studies. At the genetic level, LS513 cells express several notable features. They are positive for the p53 wild-type oncogene and express carcinoembryonic antigen (CEA) on approximately 50% of the cells. Additionally, LS513 cells express major histocompatibility complex (MHC) class I antigens, including HLA and beta 2 microglobulin, but lack MHC class II antigens (HLA-DR, DQ, and DP). The cells also produce transforming growth factor beta 1 (TGF beta-1) at a rate of 83 pg per 106 cells per 24 hours. Notably, TGF beta-1 acts as an inhibitor of LS513 cell proliferation, whereas TGF beta-2 has no significant effect on their growth. Compared to the LS1034 cell line, LS513 cells are 100-fold less sensitive to TGF beta-1, indicating distinct responses to growth factor signaling between these two colorectal carcinoma models. LS513 cells exhibit a unique profile of antigen expression, with strong positivity for intercellular adhesion molecule 1 (ICAM-1) and HLA class I antigens. The lack of MHC class II antigen expression is particularly noteworthy, as it suggests potential immune evasion mechanisms that could be relevant to colorectal cancer progression and metastasis. These features, along with their resistance to multiple drugs and their ability to form tumors in immunocompromised mice, make LS513 cells a powerful tool for studying the molecular and cellular underpinnings of colorectal cancer, especially in the context of immune interactions and therapeutic resistance. Organism: Human Tissue: Colorectal Disease: Adenocarcinoma Synonyms: LS513, LS 513 Age: 63 years Gender: Male Ethnicity: Caucasian Morphology: Epithelial-like Growth Properties: Adherent Citation: LS513 (Cytion catalog number 300457) Biosafety Level: 1 Ncbi_ Taxid: 9606.0 Cellosaurus Accession: CVCL_1386 Protein Expression: CEA+ (50%), p53+ Antigen Expression: Carcinoembryonic antigen (CEA), ICAM-1, HLA class I positive Tumorigenic: Yes, forms tumors in nude mice Products: Transforming growth factor beta 1 (TGF beta-1, 83 pg per 10 exp6 cells per 24 hours) Karyotype: Two stem lines can be distinguished. The main one was represented in 65% of the cells, with a modal number of 51,xY and 3 markers, M1 - der(1)t(1,15), M2 - der(2)t(2,3)der(3)t(2,3), M3, and a monosomy 15. The second stem line had a modal chromosome number of 52,xY and presented M2 and M3 plus an isochromosome for the long arm of chromosome 1 called M4. A trisomy 5,7, a tetrasomy 13, and a monosomy 2 and 3 were present in all of the cells analyzed, the line did not exhibit monosomy 15. Culture Medium: Ham's F12, w: 1.0 mM stable Glutamine, w: 1.0 mM Sodium pyruvate, w: 1.1 g/L NaHCO3 (Cytion article number 820600a) Supplements: Supplement the medium with 10% FBS Dissociation Reagent: Accutase Subculturing: 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. Seeding Density: 1 x 104 cells/cm2 Fluid Renewal: Every 3 days Post Thaw Recovery: After thawing, plate the cells at 5 x 104 cells/cm2 and allow the cells to recover from the freezing process and to adhere for at least 24 hours. Freeze Medium: As a cryopreservation medium, use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress. Thawing And Culturing Cells: 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. Sterility: Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods. To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections. Safety Precautions: 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. Warranty: 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. Subject To Material Transfer Agreements: If you intend to use Cytion cell lines solely for internal research at a single research site, please complete and sign our Material Transfer Agreement (MTA) and submit it along with your order.For any commercial applications - including but not limited to fee-for-service work, quality control testing, product release, diagnostic use, or regulatory studies - please complete the Intended Use Form so we can prepare a suitable agreement tailored to your project.Please note: The MTA applies only to certain cell lines. If this notice and the MTA document appear on a product page, the agreement is applicable. For cell lines not covered by the MTA, no reference to the agreement will be shown. The MTA is not valid for customers in the Americas, China, or Taiwan. Please contact our U.S. entity to receive the appropriate agreement. Required Product 1: 820600a Required Product 3: 860015.0 Required Product 4: 830100.0