Animal Cell Culture Techniques by Shirley McBride, Mary Heenan, Martin Clynes (auth.), Dr.

By Shirley McBride, Mary Heenan, Martin Clynes (auth.), Dr. Martin Clynes (eds.)

Cell tradition options let a number of molecular and phone organic inquiries to be addressed, providing physiological stipulations while averting using laboratory animals. as well as easy recommendations, a variety of specialized useful protocols masking the next components are incorporated: phone proliferation and dying, in-vitro versions for mobilephone differentiation, in-vitro versions for toxicology and pharmacology, commercial software of animal mobilephone tradition, genetic manipulation and research of human and animal cells in culture.

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For each pair of wells since they have been inoculated in duplicate. 7 it is clear that there were too many plaques to count at the lower dilutions. 19xl05 PFU. 93xl0 5 PFU/ml. 7. Example of data used to calculate plaque titer Dilution Count 1 Count 2 Total count 10- 1 10- 2 10- 3 10- 4 10- 5 10- 6 10- 7 10- 8 >100 >100 49 30 15 3 0 0 >100 >100 >200 >200 121 66 24 8 0 0 72 36 9 5 0 0 calculation 36 EDWIN O'KELLY • References Enders JF, Weller TH, Robbins FC (1949) Cultivation of the Lansing strain of poliomyelitis virus in cultures of various human embryonic tissues.

3. Thaw one aliquot of M. orale positive culture and dilute to the concentration required for the positive control. This concentration of staining for mycoplasma infection 44 MARY HEENAN, MARTIN CLYNES the mycoplasma stock should have been previously tested to ensure that the infection can be detected by the indirect test procedures in use. The procedure should be repeated for M. hyorhinis. 4. Check the coverslips microscopically to ensure that the cells have attached. Add 1 ml of conditioned medium from the test samples to each of two labeled coverslips.

Monitor the plates daily for CPE using an inverted microscope. 2 Identification of Virus Isolates Development of characteristic CPE in cell culture is often useful in making a presumptive identification of the viral isolate (Fig. 5). This identification would also be based on the specime'1 source and the cell type in which the virus has grown. However, final identification of the viral isolate needs to be confirmed . • Procedure Virus Neutralization Serial dilutions of the isolate are prepared and challenged with specific neutralizing anti-serum.

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