TCMK-1

TCMK-1 is a mouse kidney epithelial cell line that was transformed using simian virus 40 (SV40). This cell line was established to study SV40-induced cellular transformation and oncogenesis in murine kidney tissue. The transformation process resulted in cells with an altered morphology, increased proliferative capacity, and the ability to maintain continuous growth in vitro. TCMK-1 cells exhibit a predominantly epithelial phenotype with some spindle-shaped cells and multinucleated giant cells, which are common features of transformed cell lines.

One of the key characteristics of TCMK-1 is the presence of SV40 large T-antigen, which plays a role in disrupting normal cell cycle regulation by inactivating tumor suppressor proteins such as p53 and RB. This mechanism facilitates uncontrolled cell division and enhances cellular survival. Immunofluorescence studies have confirmed the expression of SV40-specific antigens in TCMK-1 cells, providing further evidence of viral-induced transformation. Additionally, despite their transformed nature, these cells have not been observed to form tumors in vivo when injected into immunocompetent mice.

TCMK-1 has been used in research involving viral oncogenesis, kidney cell physiology, and cellular transformation mechanisms. Due to its SV40 transformation, it serves as a useful model for studying viral-host interactions and the molecular pathways associated with cell immortalization. However, researchers should take into account that the presence of SV40 T-antigen may impact normal cellular functions, which could influence experimental outcomes.

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.