Vero-SF-ACF

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

1. Check all containers for leakage or breakage.
2. Remove the frozen cells from the dry ice packaging and immediately place the cells at a temperature below ­-130°C, preferably in liquid nitrogen vapor, until ready for use.

These cells may be grown in NutriVero Flex10 (Biological Industries, Cat # 05-068-1A); plus 4mM L-Glutamine (ATCC 30-2214).

The flask was seeded with cells (see specific batch information) grown and completely filled with medium at ATCC to prevent loss of cells during shipping.

1. Upon receipt visually examine the culture for macroscopic evidence of any microbial contamination. Using an inverted microscope (preferably equipped with phase-contrast optics), carefully check for any evidence of microbial contamination. Also check to determine if the majority of cells are still attached to the bottom of the flask; during shipping the cultures are sometimes handled roughly and many of the cells often detach and become suspended in the culture medium (but are still viable).
2. If the cells are still attached, aseptically remove all but 5 to 10 mL of the shipping medium. The shipping medium can be saved for reuse. Incubate the cells at 37°C in a 5% CO₂ in air atmosphere until they are ready to be subcultured.
3. If the cells are not attached, aseptically remove the entire contents of the flask and centrifuge at 125 x g for 5 to 10 minutes. Remove shipping medium and save. Resuspend the pelleted cells in 10 mL of this medium and add to 25 cm² flask. Incubate at 37°C in a 5% CO₂ in air atmosphere until cells are ready to be subcultured.

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.

Cell density: The recommended start-up seeding range is 3.0 x 10⁴ to 5.0 x 10⁴ viable cells/cm² and the recommended subculture seeding range is 2.0 x 10⁴ to 5.0 x 10⁴ viable cells/cm².

The product is provided 'AS IS' and the viability of ATCC^® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid.  Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.

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