Support Units

Cell Biology/Tissue Culture

Giuseppina Giardina
IEO, Dept. of Experimental Oncology
c/o IFOM-IEO Campus
T: +39 02 94375000
giuseppina.giardinaifom-ieo-campus.it

Members: Manuela Moia, Matteo Dal Molin, Marika Zanotti, Cristina Spinelli

Activities

GOALS
The primary goal of the Cell Biology Unit is to provide IEO researchers with healthy viable cells and other specialized cell culture techniques. Core members make available expertise, facilities, techniques and technical support required for the scientific goals of the Institute, namely the study of the cancerous cell.
Core members also work closely with researchers to develop new cell culture systems, as needed, to facilitate the scientific goals of both individual investigators and the Department as a whole.

FUNCTIONS
The Cell Biology Unit provides individual investigators with the necessary technical support, expertise, and resources for the growth of numerous cell lines. Several services are offered to help investigators place cells into culture, perform their own cell culture experiments, improve the consistency and quality of cell cultures and enhance the abilities of investigators to study biological processes in mechanistic detail. The Cell Biology Unit trains new personnel for dealing with biohazardous biological material and operates quality control testing of key biological reagents.
Moreover, the Cell Biology Unit supports investigators in the use of gene transfer technologies, including state-of-the-art viral vector technology as well as consulation with investigators to develop new recombinant viral vectors. Core members help investigators in dissection of primary normal and tumoral tissues and establishment in vitro of multiple cell populations.
The development of novel vectors, strategies of gene trasfer and growth of primary cells is an important team effort between core staff and the departmental research groups.

FACILITIES
The Cell Biology Unit accomplishes its mission by providing:
1. physical, clean space, equipment and supplies for performing cell culture work
2. cell culture services that meet the needs of the Department
3. technical assistance, education and training.

The Cell Biology Unit currently offers the following services:
-provision of space, equipment, common reagents and sterile supplies for cell culture work
-provision of training, assistance and consultation in cell culture techniques both at Biosafety level 1 and 2
-acquisition, maintenance in culture, cryopreservation and distribution to users of cell lines (please inquire for information about cell lines that are currently available through the Unit)
-testing of reagents in use for proper efficiency
-testing of cell lines for mycoplasma contamination
-testing of stably retrovirally-infected cell lines for RCR
-transfection and infection support services
Moreover, Cell Biology Unit members keep under control tissue culture protocols and books, certificates of analysis of the items tested, list of the common, departmental cell lines along with information about specific needs of the various cell lines.

GUIDELINES
The Do’s of Cell Culture Work
1. Keep all work surfaces free of clutter.
2. Correctly label reagents including flasks, medium and ampules with contents and date of preparation.
3. Only handle one cell line at a time. This common-sense point will reduce the possibility of cross contamination by mislabeling etc. It will also reduce the spread of bacteria and mycoplasma by the generation of aerosols across numerous opened media bottles and flasks in the cabinet.
4. Clean the work surfaces with a suitable disinfectant (e.g. 70% ethanol) between operations and, wherever possible, allow a minimum of 15 minutes between handling different cell lines.
5. Wherever possible maintain separate bottles of media for each cell line in cultivation.
6. Examine cultures and media daily for evidence of gross bacterial or fungal contamination. This includes medium that has been purchased commercially.
7. Keep cardboard packaging to a minimum in all cell culture areas.
The Don’ts of Cell Culture Work
1. Do not continuously use antibiotics in culture medium as this will inevitably lead to the appearance of antibiotic resistant strains and may render a cell line useless for commercial purposes.
2. Don’t allow waste to accumulate particularly within the microbiological safety cabinet or in the incubators.
3. Don't handle cells from unauthenticated sources in the main cell culture suite. They should be handled in quarantine until quality control checks are complete.
4. Avoid keeping cell lines continually in culture without returning to frozen stock.
5. Avoid cell culture becoming fully confluent. Always sub-culture at 70-80% confluency or as advised on cell culture data sheet.
6. Do not allow media to go out of date. Shelf life is only 6 weeks at +4ºC once glutamine and serum is added.