KHM-5M

The KHM-5M cell line is an important model derived from a patient with undifferentiated thyroid carcinoma complicated by neutrophilia and malignant pleurisy. This cell line is characterized by its significant production of neutrophil chemotactic factors, specifically human interleukin 8 (IL-8) and granulocyte-macrophage colony-stimulating factor (GM-CSF). These factors are crucial in the recruitment and activation of neutrophils, which play a pivotal role in the immune response and inflammation. The KHM-5M cells were shown to possess extreme chemotactic activity, a trait that was substantiated through in vitro experiments using conditioned media from the cells and the modified Boyden chamber technique.

Additionally, KHM-5M cells were transplanted into nude rats, where the infiltration of neutrophils was observed in and around the transplanted tumor tissue. This finding underscores the relevance of KHM-5M as a model for studying the interactions between tumor cells and the immune microenvironment, particularly in relation to neutrophil recruitment and function. The cell line also serves as a valuable tool for investigating the molecular mechanisms underlying cytokine production in cancer and the subsequent modification of pathological features. Through DNA cloning techniques, the chemotactic activities attributed to IL-8 and GM-CSF were confirmed, solidifying the KHM-5M cell line as a significant resource for research into cytokine-driven tumor-immune interactions.

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.