HK EGFP-Kleisin-beta

The HK EGFP-Kleisin-beta cell line represents a genetically modified variant of Hela Kyoto cells designed primarily for the study of chromosome cohesion during the cell cycle. This cell line expresses an enhanced green fluorescent protein (EGFP) fused with the Kleisin-beta protein, a crucial component of the cohesin complex that is vital for sister chromatid cohesion. The expression of EGFP-tagged Kleisin-beta allows for real-time visualization of cohesin dynamics and localization throughout the cell cycle, facilitating detailed analyses of chromosome structure and function in a cellular context.

This cell model is typically utilized in research focusing on the mechanisms of mitotic and meiotic chromosome segregation, particularly looking at how cohesin's regulation influences genetic stability and cell division. The fluorescent tagging of Kleisin-beta enables the investigation of its interaction with other cohesin components and chromosomal proteins, providing insights into the spatial and temporal assembly of cohesin on chromosomes. The use of this cell line extends to studies of genetic disorders and cancers where cohesin function is disrupted, offering a valuable tool for understanding pathogenesis and developing therapeutic strategies.

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.