Research Integrity (cell lines)

Cell lines

Validation

Only cell lines of traceble origin and identity should be used for any experiments. If you have any doubt, do consider to restock the cell line. Any cell line we use should be checked for possible contamination, from other cell lines and from mycoplasma.

New cell lines (commercial suppliers or collaborators)

  1. Check that cell lines are not included in the register of misidentified cell lines
  2. Soon after procurement, STR profile the cell lines. Check the Research Services guidelines to use the Hutchison/MRC Research Centre, which relies on the CRUK CI.
  3. Soon after procurement, cell lines should be tested to exclude mycoplasma contamination. Again, CRUK CI is kindly providing their services and lab management offers a service to liase with CI. Every cell line should be mycoplasma tested

Running cultures (every 5-10 passages)

  1. Run a fast STR profile to reconfirm the identify of the cells. This should be matched with the initial STR profile.
  2. Retest cultures for mycoplasma contamination.

Engineered cell lines

  1. Engineered cell lines have to be validated at the origin (e.g., check integration, check proper behaviour of the cells, for single cell clones keep multiple clones to assess reproducibility, use orthogonal assays to validate cell lines).
  2. STR profiling will not distinguish engineered cells from parental cells. Therfore, when handling many cultures in parallel, validate the identity of the cells by Sanger sequencing, Western Blotting or imaging,  whatever technique is the most appropriate.

No data gathered from cell lines missing an STR profile, validation or not mycoplasma tested will be considered for internal or external dissemination.

Stocking

  1. Stock early passages of newly procured cells or newly established cell lines in the main lab database and, independently, for you personally. Do not run experiments with high passages (>20-30), unless youy have established the behaviour of cell is not altered by growing in culture for extended period of times.
  2. Temporary stocking (days or few weeks) at -80C is acceptable, but cell lines should me moved as soon as possible to the core cell bank.
  3. Utilize only printed cryogenic labels (Cat. # ED1F-040WH clean template). Currently, we use DYMO LabelWriter450 and DYMO-compatible cryogenic labels from LabTag (they are pricey, so be careful not to use them for other purposes). On the side of the vessel, information should be printed both as QRcode and text. These precautions are needed to identify cells in case of damage of the labels after long-term storage. cell_lines_label_demoC- is the identifier of the Cell Lines; ZW is your user or the lab identifier, depending on database; 000 is a progressive number. Therefore, C-ZW000 should point to a record in the call bank database. XY is the user identifier of who prepared the stock and DDMMYY is the date (e.g. 311217). Therefore, XYDDMMYY should point to a labbook page reporting information about this stock and a database entry. 1M is the number of cells per vial. PN is the passage number. The QR code will hold the very same information and these are generated automatically by the DYMO Label Software. Before using it, double check it. The second label is designed with the sole scope to be user-friendly, so just print the name of the cell line (e.g. U0126-EGFP:H2B)
  4. Keep the cell bank grids up to date, including information on the STR profiles, mycoplasma testing or other validation procedures.

You will find label templates, grids and other documents on the fileserver.

Transition rules

The cut-off date for aligning to this policy was 31/12/2017 for cell line validation. Not all cell lines we have in stock should be validated, but all that are in current use shall.

The new rules for stocking, labelling and storing information will be in place immediately. The cut-off date to fully update your personal boxes is 30/06/2018