Human LOX knockout THP-1 cell line

Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Suspension

Gender

Male

Viability

~ 60%

Initial handling guidelines

Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability. <p>1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.<br>2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.<br>3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method. Based on cell count, seed cells in an appropriate cell culture flask at a density of 4x10⁵ cells/mL. Seeding density is given as a guide only and should be scaled to align with individual lab schedules.<br>4. Incubate the culture at 37°C incubator with 5% CO₂. Cultures should be monitored daily.<br>5. THP-1 cells recover slowly from cryopreservation and therefore may not be ready for subculture for a number of days. Cells should be left as much as possible over this time and only subcultured when the cell density reaches 8x10⁵ cells/mL.Small amounts of fresh media can be added until cell number/viability improves.</p>

Subculture guidelines

• All seeding densities should be based on cell counts gained by established methods.
• Cells should be seeded at 2x10⁵ - 3x10⁵ cells/mL and subcultured when they have reached 8x10⁵ cells/mL.
• It is not recommended to allow the cell density to exceed 1x10⁶ cells/mL.

Culture medium

RPMI + 10% FBS + 0.05 mM beta-mercaptoethanol

Cryopreservation medium

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.