An autoradiographic screening test for mycoplasmal contamination of mammalian cell cultures |
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Authors: | George P Studzinski John F Gierthy Jolanta J Cholon |
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Institution: | (1) Department of Pathology, Jefferson Medical College of Thomas Jefferson University, 19107 Philadelphia, Pennsylvania;(2) Present address: Medical Research Institute, Putnam Memorial Hospital, 05201 Bennington, Vermont |
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Abstract: | Summary Autoradiographic detection of incorporation of tritiated thymidine into the cytoplasm of cultured mammalian cells has been
evaluated as a test of contamination of the cultures by cell-associated microorganisms, which usually are mycoplasmas. Criteria
which indicate the presence of cell-associated mycoplasmas have been established, and the reliability of the standardized
autoradiographic method has been assessed by testing the same cultures by two colony formation methods of mycoplasmal detection.
The autoradiographic method demonstrated cell-associated microorganisms in all cultures from which characteristic colonies
were grown on mycoplasma agar. The autoradiographic method did not produce false positive results, and the outcome of this
test was evident in 3 days as opposed to 7 to 14 days by agar culture methods. Some applications of the autoradiographic method
are shown, and it is suggested that this method be employed for routine surveillance for mycoplasmal contamination in laboratories
where facilities for frequent agar culture tests are not easily available.
This research was supported by U.S. Public Health Service Grants CA 12351-02 and CA 12334-01 from the National Cancer Institute. |
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