Murine Virus Testing
Investigators must receive approval from the director of the core for a project to be initiated.
In an effort to keep the BCM mouse facilities clean of virus, the Center for Comparative Medicine has asked that cell lines injected into mice be tested for murine virus. Therefore, cell lines submitted to the core must be tested for the presence of murine virus. Cells used by this core have been tested for: Ectromelia, EDIM, Hantaan, K virus, LCMV, LDEV, MAD, mCMV, MHV, MMV, MPV, MTV, Mycoplasma sp., Polyoma, PVM, REO3, Sendai, TMEV GDVII. If an investigator uses the cells of this core rather than culture cells in his/her lab, this testing is not necessary. However, the core must test cell lines from labs at Baylor College of Medicine in order to be able to inject these cells into mice inside BCM mouse facilities particularly TMFT. The core must test all cell lines from sources external to Baylor College of Medicine such as commercial vendors or other research institutions.
Complete and submit the Murine Virus Test Request Form. Please have a purchase order number to The University of Missouri in the amount of $400 per sample to be tested as well as a charge source for an overnight shipment through Federal Express available, in order to complete the form on line.
If an investigator will have multiple clones injected by the core for his/her lab, one frozen vial of the parental embryonic stem cell used in his/her lab (i.e. AB2.2, AB2.1, etc.) must be brought to the core for testing. If the parental cells are not available, then the investigator must provide one frozen vial of cells that were electroporated and manipulated in culture in his/her lab. This will allow for testing culture contamination of Mycoplasma in that lab.
If an investigator chooses to use cells from outside the College or from a commercial vendor, all clones must be submitted for testing prior to scheduling of project.
Mouse embryonic stem cells for testing will be brought into culture on Friday afternoon.
Cells will be expanded by core personnel, into one 100mm culture dish, trypsinized and counted. Aliquots of 1 x 107 cells per milliliter will be frozen back in culture media and sent to The University of Missouri Research Animal Diagnostic Laboratory for testing.
The University of Missouri Research Animal Diagnostic Laboratory will post the results two weeks after the day samples are received in their lab.
Darwin Mouse Core personnel will contact the investigators to inform them of the results as soon as these are posted. If results come back negative the injection or electroporation will be scheduled at that time.
All samples for testing will be brought into culture on Friday afternoon.
In most cases, cells will be ready to ship the following week on Thursday. Therefore, The University of Missouri Research Animal Diagnostic Laboratory should receive the samples one week after cells are brought into culture.
Results are posted two weeks following receipt of samples.
Investigators should receive notification from core personnel approximately three weeks after sample is brought into culture.
This timeline is dependent on how fast the cells grow in culture. Therefore, cells that are slow growing or have trouble recovering from the freezing process may take additional time.
It is highly recommended that an investigator keep backup vials of cells submitted to the core. The core cannot guarantee the recovery, in culture of cell lines frozen down by other groups.
In the event that cells do not grow well in culture, the investigator will be notified as soon as core personnel become aware of the problem so that additional vials may be submitted by the investigator.
In the event the cells are positive for any of the virus tested, this core will not be able to use them in any part of the project. New cells must be generated in order to proceed with the project using the services of this core.
The director of the core will deal with other problems on a case by case basis.
After service agreement:
We would appreciate, that any publications arising from the generation of cell lines by this core carry an appropriate acknowledgment of this service.