KU812

The KU812 cell line is a human leukemic cell line originally derived from a patient with chronic myelogenous leukemia (CML) in the blastic crisis phase. It is notable for its capacity to differentiate into basophilic and erythroid lineages under specific conditions, making it a valuable tool for studying hematopoietic differentiation and related malignancies. The cell line exhibits characteristics of basophilic precursors, including the presence of metachromatic granules that are positive for toluidine blue and astra blue staining, and it synthesizes histamine, indicative of basophilic activity.

KU812 cells are particularly relevant in investigating complement activation-related pseudoallergy (CARPA) and hypersensitivity reactions mediated by basophils. This utility stems from their robust response to complement proteins like C3a and C5a, which trigger the release of histamine and other inflammatory mediators, mimicking pseudoallergic reactions. KU812 cells express cell-surface markers such as CD63 and CD203c, which are associated with basophilic activation and degranulation. These markers have been employed in flow cytometry-based protocols to evaluate the immunological compatibility of nanomedicines and other biologics.

Additionally, KU812 cells demonstrate erythroid differentiation potential when cultured in erythropoietin-supplemented conditions. This includes spontaneous maturation into erythroid cells capable of synthesizing various hemoglobins, such as adult and fetal forms. These features underline their utility in studying erythropoiesis alongside basophilic differentiation, making KU812 a versatile model for hematological research.

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.