What does the acronym SOPF stand for?
SOPF stand for Specified and Opportunistic Pathogen Free. It is also called SPF (Specific or Specified Pathogen Free). The two terms tend to be used interchangeably.
Why is SAHMRI Bioresources an SOPF-status facility?
There is a significant amount of scientific data to show that mice carrying certain pathogens react differently under certain experimental conditions. Also, there are a variety of pathogens that compromise the breeding performance and general health status of rodents.
How will my work be affected moving from a conventional facility to an SOPF one?
Generally it won't be compromised. Indeed, the reliability and reproducibility of your research will in many cases improve by moving to working under SOPF conditions. What it does mean though is that there are much stricter standards to which you and the facility must adhere. This includes the way the animals are housed, bred and handled and the clothing you wear in the facility.
What training do I require to work in the facility?
SAHMRI needs to be satisfied that you are competent with working with animals, understand your obligations and responsibilities under the Code and the State Animal Welfare Act, and are aware of how to work within the facility. The requirements are detailed here.
Can I bring equipment in to the facility?
Yes but we have strict procedures on how it can be brought in depending on what it is and where it has been housed. For example, if it is equipment that has been in another animal facility, it must first be treated using Vapourised Hydrogen Peroxide (VHP) before it can enter the facility. Contact the Bioresources Manager on how this can be arranged.
What health status do you maintain?
We adhere to the FELASA guidelines for both mice and rats in the facility. The latest guidelines are available here.
Why are mouse lines imported into the facility by embryo rederivation?
To ensure the lines we house here are clean, we must remove any pathogens they may potentially have. The only safe way to do this is by early embryo redererivation, where embryos 1-2 days old (2- to 8-cell) are transferred into clean recipients. Cesarean section is not good enough, as viral pathogens often cannot be removed by this method as they are easily transferred into the developing embryos via the placenta.
Can I bring in mouse lines from other facilities without embryo rederivation?
In the vast majority of cases, no. Even if the health status of the line appears clean the animals may still be carrying pathogens that have not been detected. The only way to ensure a clean facility is maintained is to bring everything in by embryo rederivation.
We have a couple of exceptions to this rule and these are animals that are purchased from one of the Jackson Labs clean facilities or from Charles River, USA and Charles River Europe. They must come with a detailed clean history. Such animals are screened in our Quarantine Facility upon arrival and once their pathogen-free status is confirmed they are able to be moved into the main facility.
What if I need to do a short-term experiment with a mouse lines available elsewhere but don’t want to maintain it as a line or breed here?
Unfortunately, in most cases it will still have to be embryo rederived here. We can freeze down a stock of embryos for future projects so that the line can be easily reanimated in the future when required. If the project does not involve any phenotyping, we may be able to house it for a short-period out at our Gilles Plains facility.
Do I need to have lines rederived if I am only bringing them in to freeze sperm and embryos?
Generally, no. We can bring in mice that have a relatively clean health history into our Quarantine area and freeze sperm and/or embryos there and then cull the mice. The only exceptions to this might be if the colony has a record of having contained a high-risk viral pathogen.
How do I arrange an import into the facility?
Please consult the relevant web page here for further information.
The line I wish to bring in is homozygous. Will this be altered by the rederivation process?
Yes. In almost all cases, we rederive lines by taking homozygous or heterozygous males and crossing them with our in-house C57BL/6J females. This means that the rederived GM offspring will heterozygous. Once the rederived animals are genotyped and confined clean, they will move into our breeding area where they can be intercrossed to generate the first homozygous animals.
When the line must be kept homozygous and cannot be crossed with wild type C57BL/6J for rederivation e.g. inbred lines, complex lines homozygous for several alleles we have to breed the animals up in our Quarantine are to obtain GM females to use for super ovulation. This is very expensive and avoided unless absolutely necessary.
If you do not have a genotyping assay to differentiate homozygotes form heterozygotes, you will need to develop one as we will not rederive as homozygotes for this reason only. Please consult the Bioresources Manager who can provide assistance in genotyping.
Do I need IBC approval to bring in my GM mice?
Approval by the Institutional Biosafety Committee is required but if you are only bringing in genetically modified mice that are low risk and used for breeding and simple experiments you do not need to apply for your own approval. The lines would be listed and covered under our Bioresources IBC PC1 application. Consult the Facility Manager for further details.
What stock lines do you house for researchers?
We aim to keep stocks of commonly used mouse lines for researchers that are maintained as tick-over colonies when not required so that researchers do not have to continually import a line each time they require a few animals. At the moment we have C57BL/6J and CD-1 wild type mice and CD-1 Nude mice available. We will be expanding this in the future. We also have some live and cryopreserved Flp and cre lines. Consult the Bioresources Manager for further information.
What genetic quality control methods do you use for stock lines?
We breed our wild type lines to accepted international standards that avoid genetic drift. In the case of inbred wild-type lines, we use three types of colonies (Founder, Expansion and Production) to ensure that there is a pipeline of animal breeding such that no animal can get more than 8 generations away from the original animal imported. Once, we are past 8 generations, we restock from either frozen embryos or our own cryopreserved embryos.
We also screen our lines by SNP analysis at regular intervals to determine if any contamination of the line has occurred.
Can I bring mouse tissues into the facility?
It will depend on the origin and state of the tissue. Fresh tissues from other facilities cannot be allowed in if they are in an unsealed state. We evaluate each case individually so consult the Bioresources Manager.
Can I bring mouse cell lines into the facility?
Generally, yes. We like them to be screened for mouse pathogen contamination if they are standard cell lines that are routinely cultured. We would set up a standard process (in the form of an SOP) for how each cell line is brought in and used in the facility so consult the Bioresources Manager.
Can I bring human cell lines or cells from donors or patients into the facility?
Generally, yes. We like them to be screened for Mycoplasma contamination if they are standard cell lines that are routinely cultured. We would set up a standard process (in the form of an SOP) for how each cell line is brought in and used in the facility so consult the Bioresources Manager.
How do I get my cell lines screened for pathogens?
We have an internal service run by ComPath that is able to screen cell lines for Mycoplasma and rodent viral pathogens. There are also various external services that will do this e.g. RADIL.
If the AEC has approved my project, do I need any other approvals before starting my work?
The AEC deals with the ethics around animal use and does not focus on every aspect of scientific best practice. For example, the AEC does not oversee the methods used to maintain the SOPF status of the facility. The AEC, the Bioresources manager and the Director of Research Support Services must all be satisfied your research project can be undertaken in SAHMRI. You should always consult the Bioresources Manager to ensure your SOPs are available electronically and the methods you are using adhere to the practices currently used in the facility. It is always best to show the Bioresources Manager your application before you submit it to the AEC.
What are the ARRIVE guidelines?
The ARRIVE (Animal Research: Reporting of In Vivo Experiments) guidelines were developed by the NC3Rs and are intended to improve the reporting of research using animals – maximising information published and minimising unnecessary studies.