SCLC-21H

The SCLC-21H cell line was derived from the pleural effusion of a patient with small cell lung cancer (SCLC) of the oat cell subtype. This cell line, along with SCLC-22H, was established during a period of chemotherapy, with SCLC-21H being the second to be derived after an additional 15 days of treatment. While both cell lines originated from the same patient, they display significantly different biochemical, morphological, and kinetic properties. SCLC-21H, for example, has a faster population doubling time and a higher colony-forming efficiency compared to SCLC-22H. These differences make SCLC-21H a distinct tool for studying certain variant forms of SCLC.

Biochemically, SCLC-21H differs from SCLC-22H in its low or undetectable levels of key neuroendocrine markers such as L-Dopa decarboxylase, bombesin, and carcinoembryonic antigen. However, both cell lines express high levels of neuron-specific enolase and creatine kinase isoenzyme BB, which are characteristic markers of SCLC. Moreover, while both cell lines exhibit c-myc amplification, SCLC-21H contains an additional rearranged and amplified EcoRI c-myc fragment, further highlighting its genetic uniqueness.

Structurally, SCLC-21H exhibits loose growth in culture and features prominent nucleoli and abundant cytoplasm, contrasting with the more tightly packed morphology of SCLC-22H. The presence of ultrastructurally dense core granules in SCLC-21H confirms its neuroendocrine origin, and it is classified as representing a variant form of SCLC. These distinct features make SCLC-21H a valuable model for exploring the variant forms of small cell lung cancer and understanding their response to chemotherapy.

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.