Adaptation and infection of mouse bone marrow (JLS-V9) cells in suspension culture for production of Rauscher leukemia virus.
JLS-V9 mouse bone marrow cells were readily adapted to suspension culture, chronically infected with Rauscher leukemia virus (RLV), and subsequently grown in 7.5- and 14-liter New Brunswick fermentors. The suspension-type cell system can be modified to produce virus with clearly defined properties, such as high ribonucleic acid-dependent deoxyribonucleic acid polymerase (RDDP) activity, high particle count, and high infectious particle count. Biological and biophysical properties of suspension-produced RLV were not affected by concentration and purification employing continuous-flow and rate-zonal centrifugation procedures. The RDDP assay was standardized and showed a linear incorporation of (3)H-thymidine 5'-monophosphate ((3)H-TMP) up to 30 min. Further characterization indicated that a high percentage of (3)H-TMP incorporation was due to RDDP.
D000818:Animals; D001854:Bone Marrow Cells; D002452:Cell Count; D002460:Cell Line; D002474:Cell-Free System; D002478:Cells, Cultured / Q000382:microbiology*; D002501:Centrifugation, Zonal; D004279:DNA, Viral / Q000096:biosynthesis; D007942:Leukemia, Experimental; D051379:Mice; D008854:Microscopy, Electron; D010772:Phosphotungstic Acid; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism; D011925:Rauscher Virus / Q000201:enzymology / Q000254:growth & development* / Q000302:isolation & purification / Q000472:pathogenicity; D013194:Staining and Labeling; D013942:Thymine Nucleotides / Q000378:metabolism; D014316:Tritium; D014779:Virus Replication
D004279:DNA, Viral; D013942:Thymine Nucleotides; D014316:Tritium; D010772:Phosphotungstic Acid; D012194:RNA-Directed DNA Polymerase