Working with Ciprofloxacin HCl - Cellgro

Ciprofloxacin HCl
The Best Answer to Mycoplasma Contamination Currently Available
Mediatech, Inc. 9345 Discovery Blvd. Manassas, VA 20109 800-CELLGRO (235-5476) www.cellgro.com
cellgro®
by Mediatech, Inc.
Ciprofloxacin hydrochloride (HCl) is a fluoroquinolone antibiotic. It is effective against a wide range of gram positive and gram
negative bacteria¹, and is most well known for its effectiveness against mycoplasma. Ciprofloxacin HCl works by interfering with the
bacterial enzyme DNA gyrase, an enzyme necessary for bacterial synthesis, replication, and transcription in both the active and
non-active growth phases of the bacterial life cycle2,3.
References
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Eradicates mycoplasma as
well as gram +/- bacteria
Effective at concentrations
as low as 10 μg/mL
Uninterrupted feeding
schedules
Background Information
Mycoplasma contamination may
originate from various sources including
cell culture medium, serum added to the
medium, tissue used to establish
primary cultures, and most commonly is
passed from individuals who handle the
cultures. Mycoplasma is an insidious
infection that decreases cellular activity
and growth rate4, produces membran
alterations5, affects amino acid6 and
nucleic acid metabolism, causes
chromosome aberrations7, and diminishes
the quality of resultant data or
products. Mycoplasma infections often
remain undetectable and even with the
heaviest contamination, cultures may
appear to grow normally and remain
clear. Reports estimate that up to 35% of
cultures containing mycoplasma go
undetected8. The most common
method for detecting mycoplasma is the
Barile and Kern method. If detected,
Ciprofloxacin HCl can provide the end
user with an alternative to discarding
valuable cell cultures, thus saving
valuable time and money. Ciprofloxacin
HCl shows minimal decrease in effective-
ness even after six days in culture; therefore
regular feeding schedules are not
interrupted. Treatment can be discontinued
when no signs of reoccurrence
are apparent after four consecutive
weeks9. Ciprofloxacin HCl and the protocols
for its use are patented- US patent
numbers 4,670,444 and 4,895,803.
Working Concentration
The effective concentration for Ciprofloxacin
HCl, as with many antibiotics
varies with the organism or the cell type,
enviromental conditions, and stage of
growth cycle. However, a concentration
of 10 μg/mL is active against most
strains of gram negative and gram
positive bacteria, as well as several
species of mycoplasma. In comparison
to other antibiotics, Ciprofloxacin HCl
has many advantages, including lack of
known resistant strains, minimal side
effects, a lower effective working
concentration, non-cytotoxicity, a low
reoccurrence rate for contamination,
and the ability to kill bacteria in both the
active and non-active growth phases.
Smith, B.R. “National Cooperative Study of Ciprofloxacin Susceptibility in the United States.” Current Therapeutics Research (1990) 47: 962-970.
Chalkley, L.J., and Koornholf, H.J. “Antimicrobial Activity of Ciprofloxacin Against Pseudomonas aeruginosa, Escheria coli and Staphylococcus aureus Determined
by the Killing Curve Method; Antibiotic Comparisons and Synergistic Interactions.” Antimicrob. Agents Chemother. (1985) 28: 331-342.
Wolfson, J.S., and Hooper, D.C. “The flouroquinolones: Structure, mechanisms of action and resistance, and spectra of activity in vitro.” Antimicrob. Agents
Chemother. (1985) 28: 581-586.
McGarrity, G.J., Phillips, D.M., and Vaidya, A.B. “Mycoplasmal Infection of Lymphocyte Cultures Infections with M. salivarium.” In Vitro (1980) 16: 346-356.
Wise, K.S., Cassell, G.H., and Action, R.T. “Selective Association of Murine T Lymphoblastoid Cell Surface Alloantigens with M. Hyorhinis.” PNAS (1978) 75:
4479-4483.
Stanbridge, E.J., Hayflick, L., and Perkins, F.T. “Modification of Amino Acid Concentrations Induced by Mycoplasma in Cell Cultre Medium.” Nature (1971) 232:
242-244.
Davis, J.M. Basic Cell Culture: A Practical Approach (1994) 250-251.
Hay, R.J., Macy, M.L., and Chen, T.R. “Mycoplasma Infection of Cultured Cells.” Nature (1989) 339: 487-488.
Schmitt, K., Daubener, W., Bitter-Suermann, D., Hadding, U. “A Safe and Efficient Method for the Elimination of Cell Culture Mycoplasma Using Ciprofloxacin.” J.
of Immunol. Methods (1988) 109(1): 17-25.

Working with Ciprofloxacin HCl
Mediatech, Inc. 9345 Discovery Blvd. Manassas, VA 20109 800-CELLGRO (235-5476) www.cellgro.com
cellgro®
by Mediatech, Inc.
Ciprofloxacin HCl powder should be stored at room temperature below 30°C. Liquid stock solutions should be stored frozen
(-20°C).
To prepare a stock solution add the appropriate amount of powder to distilled deionized water while mixing.
NOTE: Adding water directly to the powder may result in insoluble mixtures that cannot be filtered without losing potency.
After ensuring that the mixture is completely solubilized, filter sterilized. Store at -20°C. If desired, the pH may be adjusted.
Media containing freshly added Ciprofloxacin HCl, at an effective working concentration of 5-25 μg.mL (most commonly 10
μg/mL), should be added to cells seeded at a density of 103 to 105.
Cells should be re-seeded or sub-cultured with media containing Ciprofloxacin HCl at 3 to 4 day intervals.
Cells should be treated for 12-20 days, after which, it is no longer necesary to include Ciprofloxacin HCl in the media.
Ciprofloxacin HCl may be used in combination with other antibiotics.
Ciprofloxacin HCl is soluble in water. Stock concentrations of up to 10 mg/mL frozen can be safely recommended as far as long
term storage of liquid solution.
Product Catalog No. Size
Ciprofloxacin HCl, Powder 61-277-RF
61-277-RG
1 x 1 g
1 x 5 g