Mycoplasma Removal Agent (MRA)

Description Mycoplasma Removal Agent
Pack Size 5 ml (50 ug/ml
Product Code BUF035

Mycoplasma

Mycoplasma Removal Agent (MRA) was developed to combat the widespread problems caused by contamination of cell cultures with mycoplasma.

  • Very effective at removing mycoplasma from infected cell cultures.
  • Shows strong anti-mycoplasma activity against many types of mycoplasma including Mycoplasma orale, M. arginini, M. hyorhinis and Acholeplasma laidlawii.
  • Suitable for use to prevent recontamination of the culture with the original mycoplasma.
  • Can be used to prevent initial infection of cells in culture by mycoplasma.
  • Non toxic — will not interfere with the viability or function of cells in culture.
  • Very easy to use — simply incubate for a week after addition to mycoplasma-contaminated cell cultures.

Image legend: The effect of Mycoplasma Removal Agent (0.5 mg/ml) in the culture media of mycoplasma-infected cells after 1 week.

Basic Protocol

  1. Add MRA to cell cultures contaminated by mycoplasma at a concentration of 0.5 µg/ml and incubate for a week.
  2. For media replacement or culture transfer (passage), use a medium containing MRA at this same concentration.
  3. Transfer the cell cultures several times without MRA and confirm that regrowth of the contaminating mycoplasma has not occurred.

If mycoplasma in serum or trypsin cannot be ruled out, MRA can be added to the media at a concentration of 0.5 µg/ml to prevent contamination of cell cultures exposed to these products.

Note: The recommended MRA concentration for use is 0.5 µg/ml. The MRA concentration should be raised up to 1 µg/ml only if the recommended concentration is ineffective in removing the mycoplasma.

Cytotoxicity of MRA is low and cell toxicity is rare when used at the recommended concentration. However, it is recommended that the retention of desired cellular characteristics be confirmed after treatment.

Note: Mycoplasma Removal Agent should not be used as a substitute for good cell culture techniques!

Sample Data:

Note that the level of infection, cell type and mycoplasma strains may influence specific results. Each researcher should use the sample data as a guide from which to determine the effective MRA concentration needed with their specific cell line and mycoplasma strain.

1. Data on Mycoplasma Removal Effect – Spontaneous Infection

Concentration of MRA (ug/ml) Days*      
    0 7 14 21
Human-derived cell-A 0.39 + - - -
  0.2 + - - -
  0.1 + - + +
  0 + + + +
Human-derived cell-B 0.78 + - - -
  0.39 + - - -
  0.2 + - - -
  0.1 + + + +
  0 + + + +

(+) = Mycoplasma positive
(-) = Mycoplasma negative
(Days*) = Mycoplasma status at stage of culture (days)
Cells were incubated with MRA for 7 days.

2. Efficacy Comparison of MRA and Pharmaceuticals on the Market

  MRA   Tiamulin   Minocycline  
  MIC* MMC** MIC

 

 MMC MIC MMC
Mycoplasma orale CH-19299 0.05 0.1 0.0031 3.13 0.05 25.0
Mycoplasma arginini G-230

 

 0.1 0.2 0.0063 12.5 0.2 (>)100
Mycoplasma hyorhinis BST-7 0.05 0.1 0.0031 0.39 0.0031 0.39
Acholeplasma laidlawii PG-8> 0.0125 0.025 0.05 (>)100 0.05 (>)100
MMC/MIC 2   128 - (>) 2048   512 - (>) 2048  

()MIC = Minimum inhibitory concentration (µg/ml)
(*)MMC = Minimum mycoplasmacidal concentration (µg/ml)

References:
  1. Nakai, N. et al. (2000). Detection and elimination of contaminating microorganisms in Transplantable Tumors and Cell Lines. Exp. Anim. 49: 309 – 313.

Storage Conditions:

MRA is stable at room temperature. Protect from light to prevent decomposition.

Shelf Life:

5 years from date of despatch.