Getting Started with an ATCC Cell Line.............1
Product Sheet..............................................................1
Preparation of Medium............................................1
Initiating Frozen Cultures........................................1
Processing Flask Cultures........................................1
Cell Growth and Propagation....................................4
Passage Number and Population Doubling
Level................................................................................4
Adapting to a New Medium or Serum...............5
Temperature.................................................................5
Examination of Cultures..........................................6
Cell Counting...............................................................6
Cell Viability..................................................................7
Subculturing Monolayer Cells...............................8
Monolayer Subculturing..........................................8
Troubleshooting Monolayer Cell Subculturing...9
Suspension Cells......................................................10
Suspension Cell Subculturing.............................10
Troubleshooting Suspension Cell Subculturing...11
Adapting a Monolayer Cell Line to Grow in
Suspension................................................................11
Complete Growth Media.............................................13
Cell Culture Media...................................................13
Media Formulations...............................................14
Media Ingredients...................................................14
Media Supplements...............................................16
Osmolality..................................................................17
Antibiotics and Antimycotics..............................17
Animal Sera...............................................................18
Culture Vessels and Surfaces...................................20
Vessels.........................................................................20
Surface Coatings and Feeder Cells....................22
Cryopreservation...............................................................24
Overview....................................................................24
Freeze Medium........................................................24
Equipment.................................................................25
Liquid Nitrogen Freezer Storage.......................26
Cryopreservation Procedure...............................26
Recovery of Cryopreserved Cells.......................27
Contamination and Biosafety.................................28
Check for Microbial Contamination.................28
Mycoplasma Contamination...............................28
Treating for Microbiobial Contamination.......29
Cellular Cross-Contamination.............................29
Biosafety.....................................................................30
Glossary......................................................................................31
References................................................................................34
ATCC Media, Sera, and Reagents.........................35