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Vero-SF-ACF cell line was derived from the parental Vero cell line (ATCC CCL-81) by adaptation to serum-free and animal component-free medium. 

The parental Vero cell line has been shown to be susceptible to infection by: poliovirus 1, 2, 3; Getah; Ndumu; Pixuna; Ross River; Semliki Forest; Paramaribo; Kokobera; Modoc; Murutucu; Germiston; Guaroa; Pongola; Tacaribe; SV-5; SV40; rubeola; rubellavirus; reovirus 1, 2, 3; and simian adenoviruses.
The parental Vero cell line has been show to be resistant to infection by the following viruses: Stratford; Apeu; Caraparu; Madrid; Nepuyo; Ossa

These cells may be grown in VP-SFM (Cat. No. 11681-020, Life Technologies) according to the manufacturer's instructions.

Volumes used in this protocol are for a 75 cm² flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.

1. Remove and discard culture medium.
2. Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS).
3. Add 2.0 to 3.0 mL of 0.25% (w/v) Trypsin-0.53mM EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
   Note: To avoid clumping do not agitate the cells by tapping or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
4. Add 2.0 to 3.0 mL of 0.25% Soybean Trypsin Inhibitor and aspirate cells by gently pipetting.
5. Transfer cell suspension to a centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes.
6. Discard supernatant and resuspend cells in fresh growth medium. Add appropriate aliquots of cell suspension to new culture vessels.
7. Incubate cultures at 37°C

1. Cells grow best after the first subculture.
2. For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 13 in Culture Of Animal Cells: A Manual Of Basic Technique by R. Ian Freshney, 6th edition, published by Wiley-Liss, N.Y., 2010.

Subcultivation Ratio: 1.3 to 1:6

Yasumura Y, Kawakita Y. Studies on SV40 in tissue culture - preliminary step for cancer research in vitro. Nihon Rinsho 21: 1201-1215, 1963.

Simizu B, et al. Characterization of the Tacaribe group of arboviruses. I. Propagation and plaque assay of Tacaribe virus in a line of African green monkey kidney cells (Vero). Proc. Soc. Exp. Biol. Med. 125: 119-123, 1967. PubMed: 6027511

Rhim JS, Schell K. Cytopathic and plaque assay of rubella virus in a line of African green monkey kiency cells (Vero). Proc. Soc. Exp. Biol. Med. 125: 602-606, 1967. PubMed: 4961492

Rhim JS, et al. Temperature dependence of the synthesis of adenovirus tumor and viral antigens. Proc. Soc. Exp. Biol. Med. 127: 642-646, 1968. PubMed: 5689485

Rhim JS, Schell K. Cytopathic effects of the parainfluenza virus SV5 in Vero cells. Nature 216: 271-272, 1967. PubMed: 4293683

Rhim JS, et al. Growth of Junin virus, the etiological agent of Argentinian hemorrhagic fever, in cell cultures. Arch. Gesamte Virusforsch. 21: 243-252, 1967. PubMed: 5591575

Biosafety in Microbiological and Biomedical Laboratories 4th ed.; U.S. Department of Health and Human Services ;1999.