SK-N-LO

The SK-N-LO cell line is a human neuroblastoma cell line used in research to study neuroblastoma as well as mechanisms of apoptosis and cancer signaling pathways. It is also classified as a primitive neuroectodermal tumor (PNET) cell line and carries the EWS-FLI1 fusion gene, commonly found in Ewing's sarcoma family tumors (ESFT). This fusion gene results from a chromosomal translocation and plays a key role in the oncogenic behavior of these tumor cells.

SK-N-LO cells are particularly sensitive to certain inhibitors targeting oncogenic signaling pathways. For example, the GLI inhibitor GANT61 has been shown to induce caspase-independent apoptosis in SK-N-LO cells. GANT61 disrupts GLI1 and GLI2-mediated transcription in the Hedgehog (Hh) signaling pathway, which is critical for cell survival and proliferation in this cell line. When treated with GANT61, SK-N-LO cells exhibit morphological changes associated with apoptosis, such as chromatin condensation and nuclear fragmentation. Furthermore, GANT61 reduces the expression of proteins like GLI2 and survivin, which are important for cell cycle progression and survival, while increasing the expression of p21, a cyclin-dependent kinase inhibitor.

Additionally, SK-N-LO cells have been utilized to study opioid receptor signaling. These cells have been engineered to express the μ-opioid receptor, making them a valuable model for investigating the interaction between opioid-induced analgesia and intracellular signaling pathways. For instance, studies have shown that morphine stimulates Akt phosphorylation in SK-N-LO cells via the PI3Kγ pathway, a process that can be modulated by cAMP signaling. This highlights the versatility of SK-N-LO cells in exploring both cancer biology and neuropharmacology.

37°C, 5% CO₂, humidified atmosphere.

1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

2. 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.

3. 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.

4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

5. 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.

6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

7. 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.

8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

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.

For long-term preservation, place vials in vapor-phase liquid nitrogen at about −150 to −196 °C. Storage at −80 °C is acceptable only as a short interim step before transfer to liquid nitrogen.

Cryopreserved cell lines are shipped on dry ice in validated, insulated packaging with sufficient refrigerant to maintain approximately −78 °C throughout transit. On receipt, inspect the container immediately and transfer vials without delay to appropriate storage.

At Cytion, we stand behind the quality and viability of our products. We guarantee that our cells will be viable at the start of culture for 60 days after shipment from our facilities. We assure that all materials conform to the specifications described in the Certificate of Analysis and/or Product Sheet.

For full details, please refer to our Warranty Statement.