thawing of human cell line and starting a culture according to GLP and ISO diretions

Ensure all necessary materials and reagents are ready and at the appropriate temperature

Label all tubes and dishes with appropriate information

Thaw cell line vial in a water bath at 37°C for 2-3 minutes

Transfer contents of vial to a sterile tube and add pre-warmed growth medium

Mix gently by pipetting up and down

Centrifuge the cell suspension at low speed for 5 minutes

Carefully remove supernatant and resuspend cells in fresh growth medium

Count cells using a hemocytometer or automated cell counter

Check cell viability using a trypan blue exclusion assay

Plate cells at the desired density in appropriate culture vessels

Incubate cells at the recommended temperature and CO2 concentration

Monitor cell growth and morphology daily under a microscope

Perform regular media changes and subculturing as needed

Seed cells at the desired density in a culture dish or plate

Place culture in a humidified incubator at 37°C with 5% CO2

Monitor cell growth daily and change medium as needed

Perform regular cell passaging to maintain culture health

Ensure all equipment and supplies are properly sterilized before starting the culture

Label the culture dish or plate with the cell line name, passage number, and date

Thaw the frozen cells quickly in a water bath at 37°C

Gently add thawed cells to the culture dish or plate with fresh medium

Check for cell attachment and spreading after 24 hours

Keep detailed records of cell growth, passage number, and any changes observed in the culture