Vero (Kidney, African green monkey, Cercopithecus aethiops)

Current medium for propagation: Medium 199, 95%; FBS, 5%. The Vero cell line was initiated from the kidney of a normal adult African green monkey on March 27, 1962, by Y. Yasumura and Y. Kawakita at the Chiba University in Chiba, Japan (Nippon Rinsho 21: 1209, 1963). The cells were established in a medium consisting of 0.5% lactalbumin hydrolysate, 0.1% yeast extract and 0.1% polyvinylpyrrolidone (PVP) in Earle's BSS, 98%; calf serum (heat inactivated), 2%. The concentration of the calf serum was later increased to 5%. The cell line was brought to the Laboratory of Tropical Virology, National Institute of Allergy and Infectious Diseases, National Institutes of Health in the 93rd passage from Chiba University by B. Simizu on June 15, 1964. Beginning with the 97th passage the cells were maintained on Morgan, Morton, and Parker's medium 199, 95%; FBS (not inactivated), 5%; however, the cells have also been successfully maintained on minimum essential medium (Eagle) with non-essential amino acids and Earle's BSS, 95%; FBS, 5%. The Vero cell line has been employed extensively in virus replication studies and plaque assays. The cell line has been useful for assay of SV-40, SV-5, measles, arboviruses, reoviruses, rubella, simian adenoviruses, and polioviruses (Kitasato Arch. Exp. Med. 37: 27, 1964; Bact. Proc., p. 112, 1965; Proc. Soc. Exp. Biol. Med. 125: 119, 1967; ibid., 125: 602, 1967; ibid., 125: 636, 1967; ibid., 125: 741, 1967; ibid., 127: 642, 1968; Nature (Lond.) 216: 271, 1967; Arch. Gesamte Virusforsch. 21: 155-169, 1967; ibid., 21: 243-252, 1967). The cell line was submitted to the American Type Culture Collection in the 113th passage. DESCRIPTION OF REPOSITORY REFERENCE SEED STOCK Number of Serial Subcultures from Tissue of Origin: 121. Freeze Medium: Minimum essential medium (Eagle) with non-essential amino acids and Earle's BSS, 85%; FBS, 5%; dimethyl sulfoxide (DMSO), 10%; antibiotic-free. Viability: Approximately 85% (dye exclusion). Culture Medium: Medium 199, 95%; FBS, 5%. Growth Characteristics of Thawed Cells: An inoculum of 3 X 10(5) viable cells in 3 ml of the above culture medium per T-15 flask, yields a 30-fold increase within 7 days at 37C, provided the medium is renewed 3 times weekly and the pH is adjusted to 7.4 with a humidified mixture of 5 or 10% carbon dioxide in air. Subcultures are prepared by trypsinization. Plating Efficiency: Approximately 24% in above culture medium. Morphology: Fibroblast-like. Karyology: Chromosome Frequency Distribution 50 Cells: 2n = 60 Cells: 2 4 3 33 3 2 Chromosomes: 55 56 57 58 59 62 This is a cell line with the hypodiploid chromosome count. The modal chromosome number was 58 occurring in 66% of cells. The rate of cells with higher ploidies was 1.7%. In most cells, over 50% of the chromosomes in each cell complement belonged to structurally altered marker chromosomes. Normal A3, A4, B4, and B5 were absent; B2, B3 and B7 were occasionally paired; and B9, C1 and C5 were mostly paired. Other chromosomes were mostly present in single copy. Sterility: Tests for mycoplasma, bacteria, and fungi were negative. Species: Confirmed as monkey by immunofluorescence test. Virus Susceptibility: Susceptible to poliovirus type 3, Getah, Ndumu, Pixuna, Ross River, Semliki, Paramaribo, Kokobera, Modoc, Murutucu, Germiston, Guaroa, Pongola and Tacaribe Arboviruses. Not susceptible to Stratford, Apeu, Caraparu, Madrid, Nepuyo and Ossa Arboviruses. Reverse Transcriptase: Not detected. Submitted by: W.D. Hann(1) and J.S. Rhim(2), NIAID, NIH, Bethesda, MD. Present addresses: (1) Department of Virology, Bowling Green State University, Bowling Green, OH; (2) NCI, NIH, Bethesda, MD. Prepared and characterized by: ATCC, Rockville, MD. Price Code: J