E. Mycoplasma.
All cells being cultured in the established facility will be tested every 2 - 3 months. This will be organised by Caroline. When asked to provide a sample for testing, seed a T25 with a sample of the cell line in Pen/Strep-free medium and leave to grow for 10 days before giving the flask to Caroline
.
Bringing up cell lines from LN2 stocks that have not been tested and shown to be negative prior to being frozen down; the cells MUST be defrosted and cultured in primary culture until they have been tested and found to be negative.
Any new cell lines that are imported from external sources must be registered with Caroline. If they come from a known source (eg ATCC or ECACC) and are guaranteed mycoplasma-free then they may be cultured in established cell culture. Any cells that are not guaranteed mycoplasma-free MUST initially be cultured in primary. As soon as possible, prepare a sample for testing and freeze down a vial at the same time. once these are proven to be negative then
a new identical passage vial may be defrosted and cultured in established cell culture. We are assuming that primary culture is potentially infected with mycoplasma and therefore, UNDER NO CIRCUMSTANCES may any culture be moved from there into established cell culture.
F. Needle-stick Injuries
The UBHT policy on needle-stick injuries has changed. Any member of staff who comes into contact with a patients blood that is KNOWN to be HIV infected will now be offered HIV prophylaxis as soon as possible after the injury has occurred. Ring the Occupational Health departments needle-stick hotline on ext 4987 during office hours. Out of office hours the oncall physician should be contacted via the switchboard.
The most frequent cause for needle-stick injury is an attempt to re-sheath a needle, so NEVER RE-SHEATH A NEEDLE.
G. Decontamination Procedures
Virkon is the only anti-virally effective detergent that is used by URC-N. Therefore where decontamination from adenoviruses or human blood products is required Virkon is the only detergent that should be used for:
• routine decontamination of disposable culture ware
• routine spraying of hoods and incubators
• clearing up spills
Where cell-culture supernatants of human blood-derived cells are used in labs other than primary culture, the disposal and decontamination procedures listed must be adhered to in the lab used.
H. Radioactive Isotopes
Both communal cell culture facilities, together with the facility in lab 3, are registered for the use of tritium (proliferation assay and inositol phosphate total assay) and of S-35 (immunoprecipitation assay). Users must be registered with RPS and assigned to a radiation safety co-ordinator before using radioisotopes in any culture facility. Each user is responsible for safe usage of radioisotopes and for effective and routine monitoring. Radioisotopes should
only be used in the designated hoods and incubators. The specific trays in each facility should be used for containment. For further details, see the relevant radiation local rules for cell culture. There are NO bins designated for solid radioactive waste in ANY cell culture areas (use the appropriate bins in adjacent labs).
