CMK

The CMK cell line is a human megakaryoblastic leukemia model established from a Down syndrome patient diagnosed with acute megakaryoblastic leukemia (AMKL). These cells are significant for research on megakaryocytic differentiation and leukemia associated with trisomy 21. CMK cells display markers typical of megakaryocytes, such as glycoproteins GPIIb/IIIa and GPIb, which are key components of platelet membranes. The presence of platelet peroxidase (PPO) activity, localized to the nuclear envelope and endoplasmic reticulum but not the Golgi apparatus, confirms the megakaryocytic lineage of these cells. Ultrastructural analysis reveals the existence of cytoplasmic granules resembling α-granules and demarcation membranes, further validating their megakaryocytic nature.

CMK cells are characterized by a complex karyotype, including near-tetraploidy and a specific translocation, der(17)t(11;17), also present in the patient's original leukemic cells. This genetic alteration links the cell line back to the disease state in the patient, providing a reliable model for studying AMKL pathogenesis. Additionally, the CMK cell line can respond to hematopoietic growth factors like interleukin-3 (IL-3) and granulocyte-macrophage colony-stimulating factor (GM-CSF), which stimulate proliferation and differentiation into more mature megakaryocytic forms. Treatment with agents like phorbol ester (TPA) enhances the expression of megakaryocytic markers and induces morphological changes, including cytoplasmic segmentation resembling platelet formation. These features make CMK a valuable tool for investigating megakaryocytic lineage development, the effects of trisomy 21 on hematopoiesis, and the mechanisms of leukemogenesis in Down syndrome.

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.