This unit was established in Iranian Tissue Bank in 2004 for human or animal cell culture. As a result, cell culture has become a central technique in current molecular and cell biological research. We have clean room facility in ITB and all of our biological activities for clinical applications is done in clean room environment and upon Good Cell Culture Practice (GCCP) and other cell and tissue banking standards.
The cell culture facility provides technical supports. our facilities consists of laboratory for cell culture procedures, a class 10000 clean room including one separate room for instrumentation and preparation and sterile filtration of media and other facilities contains:
1- three class 100 laminar flow hoods
2- three -196oc liquid nitrogen freezers
3- two controlled rate freezers
4- two CO2 incubators
5- two refrigerated centrifuge
6- two inverted microscopes
7- three stereomicroscopes
8- one PH-meter
9- one water purification system
10- one steam sterilizer
11- one oven
12- one PCR system
13- one flow cytometer
14- one -20oc mechanical freezer
15- two -86oc mechanical freezers
Cell Culture Steps
1- Cell Isolation
Cells can be isolated from tissues for ex vivo culture in several ways. Cells can be easily purified from blood or cells can be released from soft tissues by enzymatic digestion with enzymes such as collagenase, Dispase, with or without hyaluronidase, which break down the extra cellular matrix such as Schwann cell Isolation from peripheral nerve(ITB method). Alternatively, pieces of tissue can be placed in growth media, and the cells that grow out are available for culture.
2- Culture of Isolated Cells
Isolated cells are putted in plates or culture flasks with culture medium and other cell culture additives as factors at an appropriate temperature and gas mixture (typically, 37°C, 5% CO2) in a cell incubator that this conditions vary for each cell type. All manipulations are typically carried out in laminar flow cabinet to exclude contaminating micro-organisms. Antibiotics can also be added to the growth media (usually Penicillin/Streptomycin).The common manipulations in cell culture are media changes, passaging cells, and harvesting cells.
3-Change of Culture Medium
The purpose of media changes is to replenish nutrients and avoid the build up of potentially harmful metabolic byproducts and dead cells. In the case of suspension cultures, cells can be separated from the media by centrifugation and resuspended in fresh media. In the case of adherent cultures, the media can be removed directly by aspiration and replaced (such as Schwann cell).
Suspension cultures are passaged with a small amount of culture containing cells putted in a larger volume of fresh media. For adherent cultures, cells first need to be harvested and a small number of harvested cells can then be used to seed a new culture.
5- Cell Harvesting
Tissue culture and Tissue engineering
Cell culture is a main component of tissue culture and engineering. In tissue engineering, cells are often grown and maintained in a standard culture until they are seeded into a engineered scaffold. The considerations of cell culture persist during tissue culturing, often with added complexity as tissue cultures tend to adopt more three-dimensional form, more extensive maintenance and directive stimuli to induce functionality as a tissue.