LABORATORY PROCEDURES FOR ANIMAL & HUMAN CELL LINES
REFERENCE NO: AHC/1998/4/2.2 Appendix 4
TITLE: HELPFUL HINTS - ECACC -
PROCEDURES FOR HANDLING CELL LINES UPON RECEIPT
Transfer ampoule(s) to a storage vessel if it is not to be used immediately. Cells should be stored in the gaseous phase of liquid nitrogen if not to be used immediately.
Note: This may result in some loss of viability.
On arrival check the cells under an inverted microscope and incubate overnight.
Please contact ECACC within 24 hours if there appears to be a problem.
A. ADHERENT CELLS
In order to split the cells they should be detached from the flask by trypsinisation as follows:
i) wash the cells twice with PBS
ii) add the appropriate amount of trypsin/EDTA solution (1ml per 25cm²), swirl to ensure all the surface is covered, then decant most of the solution leaving just sufficient to wet the cells.
B. SUSPENSION CELL LINES
- CELLS THAT GROW AS CLUSTERS: cells that normally form clusters should be allowed to settle, the exhausted medium pipetted off and the cells resuspended in fresh medium as centrifugation may cause n cell damage.
- SEMI-ADHERENT CELL LINES : certain cell lines grow in suspension and also attach to the tissue culture flask. Knock the flask gently to dislodge the cells and treat as a suspension cell line.
Guidelines prepared for CABRI by CERDIC, DSMZ, ECACC, INRC, November 1998
© The CABRI Consortium 1999-2013.