Efficiency of human rotavirus propagation in cell culture

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Abstract
This study was designed to find methods to reproducibly propagate human rotaviruses from fecal specimens and to determine the relationship between particle numbers and infectivity. Growth of virus was initially compared in primary and continuous lines of monkey kidney cells. Primary cells (African green and cynomolgus monkey kidney) supported virus growth directly from fecal specimens much more efficiently than did continuous lines of African green (CV-1) or rhesus (MA104) monkey kidney cells. Rotaviruses were grown in primary cells from 14 of 14 fecal specimens of different individuals collected over 3 yr. Although rotaviruses in fecal samples could not always be grown in the continuous cell lines, 2 passages in primary cells appeared to fully adapt the viruses for propagation in the continuous cell line tested (MA104). The efficiency of rotavirus growth was quantified with 5 of the fecal isolates. On the average, 1 of every 46,000 particles in fecal specimens infected monkey kidney cells. After 3 passages in primary cells, an average of 1 of every 6600 progeny virus particles appeared to be infectious. Thus, rotaviruses in fecal specimens were consistently grown in primary cells; passage in these cells increased virus infectivity and adapted the viruses for growth in continuous cell lines.