Characterization of porcine stable kidney cell line adapted to hyperthermic temperature.
The optimum temperature for the growth of porcine stable (PS) kidney cell line is 37 degrees C. We have adapted the cell line to grow at 40 degrees C. The original cell line grown at 37 degrees C has been denoted as PS-37, and the adapted new strain has been denoted as PS-40. Both the cell lines were screened for mycoplasma by Hoechst staining and tritiated uridine-uracil uptake and were found to be negative. Comparative characterization of PS-40 and its progenitor PS-37 cell line was done by using various parameters. The antigenic studies indicated that the new cell strain was not cross-contaminated with any other cell lines. It was observed that PS-40 cells were more fibroblastic with clean cytoplasm and appeared healthy. The growth of PS-40 cells was faster than the original cell line. The karyological study showed heteroploid chromosome number in PS-40 cells. The modal chromosome number of PS-40 cells was 58, whereas that of PS-37 cell line was 38. The lactic dehydrogenase isoenzyme pattern showed a cathodal shift of bands. The PS-40 cell strain could be cryopreserved and revived. The viability of PS-37 as well as PS-40 cell lines is in the range of 90-95%, and the growth characteristics of thawed cells showed six- to eightfold multiplications within 5 d. The virus susceptibility study revealed that the cytopathic effect was more profound and observed 1 d earlier in PS-40 cell line. Increased yields of Japanese encephalitis, Sindbis, and Semliki forest viruses were obtained by 1.8, 1.75, and 1.5 log plaque-forming units/ml, respectively. The yield of West Nile virus was, however, comparable to that in PS-37 cell line. Both the cell lines were refractory to Dengue viruses.
Paranjape
Smita
S
National Institute of Virology, 20A Dr. Ambedkar Road, Pune 411 001, Maharashtra, India. spparanjape@hotmail.com
Patil
B R
BR
Kadam
V D
VD
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D002460:Cell Line; D002470:Cell Survival / Q000502:physiology; D003588:Cytopathogenic Effect, Viral / Q000502:physiology; D007527:Isoenzymes / Q000378:metabolism; D007621:Karyotyping; D007668:Kidney / Q000166:cytology*; D007770:L-Lactate Dehydrogenase / Q000378:metabolism*; D034421:Sus scrofa; D013696:Temperature; D014773:Virology / Q000379:methods; D014779:Virus Replication
D007527:Isoenzymes; D007770:L-Lactate Dehydrogenase