Culturing MA-104 Cells for Rotavirus Propagation

MA-104 cells (African rhesus monkey kidney) were purchased from the ATCC CRL-2373.1 and maintained in Eagle’s minimum essential medium (EMEM) supplemented with 5% FBS, 100 µg/mL streptomycin, 100 U/mL penicillin, and 100 U/mL amphotericin B [20 (link)]. The 85A (porcine rotavirus, G5P[7]) and KJ-56 (bovine rotavirus, G8P[7]) rotaviruses were isolated from fecal samples of diarrheic Korean piglets and calves. The rotaviruses were pre-activated with 10 µg/mL trypsin (1:250) procured from the GIBCO Invitrogen Corporation, California for 30 min at 37 °C before inoculation into MA-104 cells (confluent) [10 (link)]. Afterward, the infected MA-104 cells were cultured in a 1 µg/mL trypsin-containing medium.