HK EGFP-LaminA/H2B-mCherry

The HK EGFP-LaminA/H2B-mCherry cell line is a genetically engineered Hela Kyoto derived cell model developed to facilitate advanced studies in nuclear dynamics and chromatin organization within living cells. This cell line expresses two fusion proteins: EGFP (enhanced green fluorescent protein) fused with Lamin A, and mCherry (a red fluorescent protein) fused with Histone H2B. The EGFP-Lamin A fusion highlights the nuclear envelope and allows for the visualization of nuclear architecture changes during cell cycle progression or under various experimental conditions. Meanwhile, the H2B-mCherry fusion protein binds to DNA and provides a vivid red fluorescence that marks chromatin, enabling real-time observation of chromosomal processes during mitosis and interphase.

These cells are invaluable for real-time imaging applications, including studies on nuclear integrity, DNA replication, and cellular aging, as well as research into diseases where nuclear architecture is disrupted, such as cancer and laminopathies. The dual-color fluorescence feature of this cell line allows for simultaneous visualization of both the nuclear envelope and chromatin, facilitating a comprehensive understanding of nuclear-cytoplasmic interactions and the spatiotemporal organization of chromatin. Such capabilities make it a critical tool for molecular biology research and cellular biophysics, providing insights into the mechanics of gene expression regulation, nuclear organization, and the cell cycle.

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.