OK Cells

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.

The OK cell line is a permanent epithelial-like cell culture derived from kidney tissue of an adult female American opossum (Didelphis virginiana). Established in vitro, this cell line is notable for its non-diploid chromosomal modal number of 23 and its adaptability to tissue culture conditions. Initially derived from mixed cell types, the culture evolved into predominantly epithelial cells after eight passages. The OK cell line has been extensively characterized in terms of morphology, chromosomal constitution, and growth dynamics, making it a robust model for cytogenetic and chromosome isolation studies.

One of the key features of the OK cell line is its utility in chromosome studies, especially for isolating the mammalian X chromosome. The opossum X chromosome is significantly smaller (approximately 30% smaller than the smallest autosomes) and does not contain large blocks of constitutive heterochromatin, facilitating separation from autosomes through techniques like flow microfluorometry and gradient centrifugation. The stable karyotype of the OK cells, with the presence of a distinctive metacentric marker chromosome, enhances their application in genomic and chromosomal studies. The paternal X chromosome's preferential inactivation in this marsupial provides a comparative model for studying mechanisms underlying X-chromosome inactivation in mammals.

OK cells have also demonstrated resilience and adaptability in various culture conditions, including serum variations and different mitotic-arresting agents like Velban (vinblastine sulfate), which is particularly effective for achieving high mitotic indices for chromosome isolation. The cell line's ability to synchronize and produce high yields of metaphase cells further underscores its suitability for detailed chromosomal analyses, including DNA content quantification and high-resolution imaging of chromosomal spreads.

Monolayer, adherent

Female

1

Supplement the medium with 10% FBS and 1% NEAA

2 to 3 times per week

Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

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.

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.

Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

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.

Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

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.

Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

When planning to store a cryovial in liquid nitrogen for future thawing, it is mandatory to adhere to stringent safety measures. Appropriate protective gloves and clothing are essential, and the use of a face mask or safety goggles is required during the transfer of frozen samples to or from the liquid nitrogen tank. This is to mitigate the risk of injury from potential cryovial explosions upon removal, which can result in the projection of sharp fragments.

We stand by the promise of delivering products with high cell viability and robust culture performance. To achieve the best results, please make sure you follow the storage and culture instructions detailed in the product information sheet closely. Your adherence to these guidelines is key to success.

Alpha2-adrenergic, serotonin, parathyroid hormone, atrial natriuretic factor

Accutase

CLS-820100a, CLS-860015, CLS-830100, CLS-831100