SKW-3

The SKW-3 cell line, originally believed to be derived from the peripheral blood of a 61-year-old male diagnosed with chronic lymphocytic leukemia (CLL), represents a significant point of interest in cancer research, particularly in the study of B-cell leukemias. Over time, critical reassessments using Short Tandem Repeat (STR) profiling have illuminated an important issue-SKW-3 cells are not a pure line from the CLL patient, but are instead contaminated, now identified as a derivative of the KE-37 cell line. This revelation has profound implications for past research and future studies, emphasizing the need for rigorous cell line authentication to ensure experimental accuracy.

KE-37, the true origin of the SKW-3 cells, is a B-cell line established from a patient with acute lymphoblastic leukemia (ALL). This shift in origin from CLL to ALL, due to the contamination, drastically alters the biological context and utility of the SKW-3 line. For researchers, this means that any findings or data previously attributed to CLL-specific mechanisms when using SKW-3 must be critically evaluated and potentially revised. The reclassification to a derivative of KE-37 necessitates a shift in the application of SKW-3 cells towards studies more relevant to ALL and its underlying mechanisms, rather than CLL.

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.