Team:XHD-WS-Wuhan-A/experiment/safety/
Safety
Safety Training
The iGEM official site has given very detailed guidelines and directions on how to safely proceed when doing biological experiments and behave when in a biological laboratories. Before we were allowed to enter the laboratory, we were given a speech of the safety in laboratory and a population of biography knowledge by our professor. He informed us of every potential risks and dangerous associated with the equipment and tectonics in the coming experiments, as well as how to use these tools, how to minimize the damage and how to avoid or deal with these risks. We also underwent some professional training conducted by credited teachers on safety precautions and laboratory skills before we started the sections in our project. And then, we were given a laboratory induction from our professor, and she told us the most efficient and safest way to carry out the laboratory protocol we would require.
In our experiments, many steps like transfection, which associate with the live organism or cells, should be done in aseptic surrounding, so the procedure like this will be operated in the Biosafety Cabinet, including the treatment of the gastric cancer cells. In addition, precaution for handling and disposal of waste liquid and container were also carried out according to the standard operating procedure.
Safety
Some of the laboratory equipment we use, especially glassware, can become dangerous if ruptured, and some of the reagents we use are skin and/or eye irritants. When using molecular cloning of E. coli, there is a risk of infection of ourselves and contamination of the equipment we use, which may lead to some infection on others. In order to avoid the occurrence of such accidents as far as possible and to minimize the damage after the accident, for the preparation, we received general biosafety laboratory safety training, hazardous waste management and general chemical hygiene from the researchers from the Office of Environmental Health and Safety of Wuhan University's School of Basic Medicine. To ensure the safety of ourselves and the biological materials we use, we use appropriate personal protective equipment, including gloves, lab clothes and, if necessary, glasses and so on. To avoid contamination, we collect toxic waste separately and timely submit it to the school's biological hazardous chemicals treatment center for unified treatment.
Bio parts safety
When using cell lines, including the human cell lines we used in our experiments, GES-1, MGC-803, and SGC-7901, the risk group of these cell lines comes from NH1, the risk class is RG1, is harmless to humans, and is environmentally harmless, too. In order to remain these cell lines sterile, our experiments are carried out in a separated and closed room in the laboratory, and all mammalian cell experiments are conducted in biosafety cabinets and experimental operators are worn in laboratory coats and shoe covers that are different from those we use when dealing with common bacteria.
In our experiment stoic-test miRNA sensors, we co-transfected RNA over-expression plasmids with miRNA targets to mammalian cells, which are safe for people and do not involve ethical risks. And our personal protective equipment is sufficient to protect us from other harms, as well as our transfection systems are neither contagious nor self-replicating, reducing the safety risks of using these materials.
Engineering design safety
To further improve the safety coefficient of our project, we decided to design a diagnostic method which is extracorporeal rather than esoteric, eliminating patients’ concerns that the program may have any health-related side effects. In addition, since our project is diagnostic, we decided to produce a non-toxic green fluorescent protein and a red fluorescent protein for positive diagnosis. We are interested in developing a new method for diagnosing stomach cancer in the future, which will establish four miRNAs sensors to detect the expression of miRNAs in different human gastric cancer cell lines and normal gastric cell lines with a miRNA over-expression plasmid. These miRNAs were selected because one of them expressed higher in patients’ cells with stomach cancer and the other one expressed higher in patients’ cells with early gastric cancer. We will test the potential role of plasmids in gastric cancer cells to detect miRNA expression, and plan to develop new non-invasive gastric cancer diagnoses in the future based on detecting the expression of specific miRNA. We will conduct experiments in the professor's lab where cell lines, bacteria (E. coli DH5 alpha) are not at risk to us, our communities, and our environment. Recombinant enzymes are inherently dangerous because they can reverse DNA in one direction, but since the plasmids of the encoded recombinant enzyme are introduced into the cells of live tissue biopsy samples, not in the living cells, it will not harm any patients who will receive our diagnosis at all.
Safety Goals
1. When designing the experiment, assess the risk of all reagents, microorganisms, and the substance they may produce, and design protective measures beforehand.
2. When experimenting, use standard lab safety techniques to reduce risk of physical or chemical danger.
3. Any substance or organism used shouldn’t cause disease in any individual involved.
4. Any pathogen involved shouldn’t spread outside the laboratory nor infect the community.
5. The experiment should not contaminate the environment in any way.
6. The experiment will not cause any unnecessary waste of experimental drugs or equipment damage, saving is Paramount.
7. Meet all iGEM’s safety and security rules along with all terms mentioned above.
Lab Protocols
1. There must be professional supervision during all parts of the experiment.
2. Standard precautions are to be followed at all times.
3. All experimenters should wear lab coats and gloves before entering the lab.
4. After entering the laboratory, all experimenters are not allowed to entertain, eat or use cosmetics, and keep the environment clean and tidy.
5. The drug used in the experiment should be returned to the original place after using.
6. The waste liquid and waste residue which was used in the experiment should be collected, discharged to some designated location according to regulations. And don’t to dump waste liquid and waste solvent into the sewer.
7. Laboratory equipment and medicines should be cleaned immediately after use.
8. After the experiment, experimenters should clean up the waste-bin, disposal bottle and sink pipes to prevent blockage and corrosion.
9. Everyone should pay attention to the use of flammable, explosive hazardous and highly toxic substances to prevent fire or unnecessary damage.
10. Before leaving the laboratory, all experimenters will be forced to wash their hands with 70% alcohol to ensure maximum safety.
11. Before leaving the laboratory, experimenters should ensure that the water, electricity, doors and windows are closed properly, and ensure that the laboratory equipment are closed properly to avoid safety and hygiene problems or property damage.
12. Pay attention to wearing protective equipment and clothing when using toxic drugs to prevent poisoning.
13. The experimental operation should be carried out in a sterile environment to avoid infection of experimental drugs and products affecting the results of the experiment or causing safety risks.
14. When using ultraviolet equipment, experimenters should pay attention to protect themselves, wear protective clothing and goggles in order to avoid uv damage to the skin or eyes.
15. All experimenters involved in performing the experiment must be familiar with the laboratory operating procedures and safety measures before they start the experiment.
16. All experimenters involved should be familiar with the inhalation and injection risks before doing experiment.
17. All experimenters involved must be familiar with the standard procedures of dealing with spillage onto work surface or skin, or other accidents.
18. All experimenters involved must be familiar with the correct disposal and decontamination methods of toxic material.
Facility Security
We have access to biological safety cabinets and laminar flow cabinets to ensure the safety of our experiment. The Laboratory of the Department of Basic Medical Sciences of Wuhan University, located next to the Zhongnan Hospital, is helping our experiment. The laboratory is equipped with complete exhaust ventilation ducts, eye washer, emergency spray level, fire extinguisher, fire blanket and other safety protective equipment. All the equipments meet the medical laboratory standards. And we will be guided by professionals to ensure the safety of the experiment to those performing it and the community and environment around it.
Further Safety Consideration
We have decided to use E. coli which is nonpathogenic and will have a hard time reproducing on its own in natural environment. The strain used will remain within the laboratory during the experiment. After the experiment is finished, all cultures containing, or have been in contact with, the strain will be sterilized by high temperature and pressure. All the reagents used would not cause pollution in the environment. All potentially contaminating waste is to be disposed of properly, and would be treated with unified and professional procedures. As we move through the experiments, shipment security and potential further contamination has been considered and assessed to ensure that the final project would not be infectious or contaminate the environment.
On the top of that, we also intend to use gastric cancer organelles from patients. It is difficult for the organelles to survive, to reproduce in the natural environment and to cause potential damage and impact on public or environment.
Address
XHD-WS-Wuhan-A
Wuhan High School
No.538,Gongxiu Road,Xiaoshan District,
Wuhan,Hubei Province,China