HT-3

HT-3 is a human cervical cancer cell line derived from squamous cell carcinoma and is noted for being HPV-negative. This cell line has been widely used in studies focused on cellular physiology, particularly in relation to ion channel regulation, signal transduction, and the role of cytoskeletal dynamics in cellular processes. HT-3 cells demonstrate activation of volume-sensitive chloride (Cl⁻) channels, which are crucial for their regulatory volume decrease (RVD) responses under osmotic stress. This mechanism is pivotal for maintaining cellular homeostasis by allowing the efflux of Cl⁻ and K⁺ ions, thereby facilitating water movement out of the cell to restore normal volume after swelling.

Research into the HT-3 cell line has highlighted the involvement of specific protein kinase C (PKC) isoforms in volume regulation. Notably, PKC-α has been implicated in the activation of volume-sensitive Cl⁻ channels and RVD responses. Experimental data show that inhibition of PKC-α, using either chemical blockers or isoform-specific antibodies, significantly impairs these channels' function, leading to prolonged cell swelling and disrupted RVD. This underscores the role of PKC-α as a regulatory factor in HT-3 cells' response to hypotonic stress.

Additionally, the integrity of the cytoskeleton, particularly actin filaments and microtubules, has been demonstrated to be essential for the proper activation of Cl⁻ channels and effective volume regulation in HT-3 cells. Disruption of actin filaments with agents such as cytochalasin B enhances Cl⁻ conductance and affects cell volume recovery, while stabilizing agents like phalloidin inhibit these channels' activity and delay RVD. Microtubule-modulating compounds, such as colchicine and taxol, have differential impacts, highlighting the complex interplay between cytoskeletal structures and ion channel function in these cancer cells.

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.