Tissue Culture Series #1: How to Thaw Cells with High Efficiency

Tutorial demonstrating best practices for starting a fresh cell culture from frozen stocks, including cell thawing tips and tricks to maximize survival.

• Ice crystals and thaw time
• How to avoid osmotic shock
• Cryoprotectant removal and media replacement
• Thawing stem cells and primary cells
• Methods for resuspending cells
• Seeding density considerations
• Troubleshooting

[misscarebearclaire]

I was told that if you didnt do a second spin down/resuspension that there would be remaining DMSO left over and cause cell damage. Is this true? or is that extra spin causing unneccessary stress

[funny]

Regarding using the antibiotics for primary cells : Does is it necessary to add antibiotics whenever we change the media( let us say every 24 hours) for the first passage P0, or it is recommended to add antibiotics just for the first 24 hours ?

[SueOwais]

I could not understand what exactly happens after aspirating cells. Please explain.