Bardet-Biedl Syndrome (BBS) is a rare genetic disorder with highly variable symptoms which may include retinal degeneration. BBS has a low prevalence in the population, affecting less than 1 in 100,000 people. Patients with BBS can experience problems with obesity, specifically with fat deposition along the abdomen, and they often suffer from intellectual impairments.
NCI-H358 is a human non-small cell lung cancer (NSCLC) cell line that is widely used in research related to KRAS gene mutations. The KRAS G12C mutation is relatively common in NSCLC and the NCI-H358 cell line harbors this mutation, which plays a critical role in tumor cell signaling and proliferation. Tumor cells carrying the KRAS G12C mutation may exhibit sensitivity to specific KRAS inhibitors. Because of this, NCI-H358 is invaluable for drug screening, particularly for testing KRAS inhibitors like Sotorasib (AMG 510), which targets the KRAS G12C mutation by forming a covalent bond with the cysteine residue at position 12 of the KRAS mutant protein, inhibiting its activity and affecting downstream signaling.
Cell Name: NCI-H358 (Human Non-Small Cell Lung Cancer Cell)
Catalog Number: TCHu151 (Chinese Academy of Sciences)
Growth Characteristics: Epithelial-like morphology, adherent growth
Culture Conditions: 1640 medium + 10% FBS
Culture Environment: 95% air + 5% CO₂; 37°C
NCI-H358 cells are essential in various lung cancer research applications, including:
- Drug Screening: NCI-H358 cells exhibit sensitivity to various anticancer drugs, making them an ideal model for evaluating the efficacy of new drugs, especially those targeting the KRAS G12C mutation.
- Gene Function Studies: Researchers use NCI-H358 to study the roles of specific genes in lung cancer by employing gene knockout or overexpression experiments.
- Signal Pathway Analysis: This cell line is used to investigate intracellular signaling networks, particularly those related to the KRAS pathway.
- Tumor Microenvironment Simulation: NCI-H358 is used to study interactions between tumor cells and surrounding cells, such as immune cells and stromal cells, providing insights into the tumor microenvironment.
- Add 6 mL of complete culture medium to a centrifuge tube to begin the thawing process.
- Thaw the cells in a water bath until the ice block is the size of a rice grain, then stop the water bath.
- Transfer the cells to the centrifuge tube and centrifuge.
- Resuspend the cells and seed them in a suitably sized culture dish.
- After 24 hours, observe the cell adhesion and change the medium once.
- Aspirate the supernatant and rinse the cells with room temperature PBS once.
- Add trypsin, ensuring it evenly covers the bottom of the dish.
- Digest the cells until some start to detach when tapping the dish, then stop the digestion.
- Gently pipette the cells to disperse them and transfer them to a centrifuge tube for centrifugation.
- Resuspend the cells and seed them at the appropriate ratio.
- Digest and centrifuge the cells to obtain a cell pellet.
- Resuspend the cells in cryopreservation solution and transfer them to cryovials.
- Place the cryovials in a pre-cooled (4°C) controlled-rate freezing container, then store them in a -80°C freezer overnight.
- Transfer the cryovials to liquid nitrogen for long-term storage the next day.
- Serum Quality: Use high-quality fetal bovine serum; consider increasing the concentration slightly.
- Digestion Monitoring: Avoid over-digestion by closely monitoring the process.
- Medium Replacement: As cell density increases, replace the medium or passage cells regularly to maintain optimal growth conditions.
Cyagen offers comprehensive gene editing services for the NCI-H358 cell line using the Smart-Targeted Gene Editing™ system, ensuring precise and efficient genetic modifications.
Gene modifications in the NCI-H358 cell line include gene knockout, point mutation, knock-in (KI), overexpression, and interference. Cyagen optimizes the culture conditions and monoclonal preparation for NCI-H358, enabling the delivery of NCI-H358 gene-edited monoclonal cells. This significantly enhances the accuracy and reliability of experiments, providing a more solid foundation for tumor immunology research.
- Ensure that the cells are in good condition before transfection, with a cell density of around 70%.
- During digestion, try to disperse the cells into single cells as much as possible.
- Use the limiting dilution method to prepare monoclonal cells, employing Cyagen's custom culture medium and adding an additional 10% serum. Once clones are observed, supplement with fresh culture medium and continue cultivation.
The NCI-H358 cell line is a powerful tool in lung cancer research, particularly for studying the KRAS G12C mutation. By following best practices for cell culture and gene editing, researchers can unlock new insights into cancer biology and therapeutic strategies. Cyagen’s expertise in gene editing further enhances the research potential of NCI-H358 cells, offering precise and reliable modifications that support groundbreaking studies.
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Contact our experts and take advantage of our optimized Targeted Gene Editing-Pro gene edited cell line modeling service platform for low-cost cell line generation with rapid turnaround and stable expression.
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