BeAeonTM retards the senescence of MRC-5 cells

It is well known that telomerase is the main agent to protect telomere during the cellular division [1].Recent tests, in particular on mice, showed the efficiency of certain plants whilst determining that these plants had nocarcinogenic effect [1, 2].

The Nobel Prize of Physiology or Medicine 2009 was awarded to Elizabeth H. Blackburn, Carol W. Greider and Jack W. Szostak for their discovery of the protection of telomeres by the telomerase.

We tested plant extracts of astragalus on fibroblast human cells of type MRC5 and found evidence of a meaningful effect on the extent of human cell life [3].

Experiments were carried out on a MRC5 cell strain fed with a DMEM medium (Dulbecco's Modified Eagle's Medium) and a «telomerase booster » the compounds of astragalus root.

MRC-5 cells are a human diploid fibroblast culture able to duplicate in vitro for approximately 50 population doublings, after which they become senescent and cease to replicate i.e. they are a ‘finite cell ine’.

We worked on the strain labeled 11D007 dated 04/2011 and aged 30 population doublings.

Protocol

The medium is made from DMEM medium and is mixed with these additional compounds:

    • fetal calf serum 10%
    • Glutamine 1%
    • Penicillin (10,000 U/ml) & Streptomycin (10,000 µg/ml) 1%

The MRC5 sham samples and the MRC5 fed with our « telomerase booster » (TB for short) were grown in wells under usual growing conditions (CO2, Temperature,...).

During a first experiment we got 6.4% additional cells for MRC5 fed with our TB.

After adjusting the percentage of compounds within the TB, we launched a second experiment and achieved a cell increase of 19% when fed with our TB(BeAeonTM).

To determine whether the difference between the amount of additional cells in the Sham sample and the modified medium resulted from senescent cells still active rather than young divided cells, we engaged a new experiment. This experiment's main characteristic lies in the limited space factor for cells to expand for ever. This limit on cell expansion blocks the number of cells to a global extent in such a way that all the samples achieve the same number of cells. The optical density activity level of these samples is in direct proportion to the age of MRC5 cells since cell activity declines as cells age.

To get a global figure on the activity level of our samples we worked with the XTT test.

Samples were grown in two independent series of 12 independent wells for a week.

In order to balance the noise figure linked to some cell amount differences, which were compared to each other regarding their optical activity level, we made the assumption that cell samples could reach a random difference of up to 20% in amount. Generally we didn't see such a variation in levels but we took into account this noise figure in order to discard any outlying results.

Results

As cells age the death rate settles as a down slope activity under the XTT test.

Given that the death rate of the sample fed without our TB is settled as «1» we got an half death rate for our samples fed with our TB.

Given that the MRC5 sample fed with our TB is populated with as many cells as in the MRC5 sham sample we achieved 2 times more «surviving» cells in our TB samples.

This leads to a conclusion that these surviving cells are «senescent» cells still functioning at an acceptable level of activity according to the XTT test.

As a global conclusion, linked to our previous experiments and data gathered from scientific literature, our TB helps cells to preserve the telomere caps so that cells could survive a longer time than natural cells usually do.

According to our biological model and the noise range as specified above the additional life expectancy under a TB prescription could be 1.6 time higher than the normal life expectancy.

1. Bernardes de Jesus, B., Vera, E., Schneeberger, K., Tejera, A. M., Ayuso, E., Bosch, F. and Blasco, M. A. (2012), Telomerase gene therapy in adult and old mice delays aging and increases longevity without increasing cancer. EMBO Mol Med. doi: 10.1002/emmm.201200245.
2. Calvin B. Harley,1,6 Weimin Liu,2 Maria Blasco,3 Elsa Vera,3 William H. Andrews,4 Laura A. Briggs,4 and Joseph M. Raffaele5. REJUVENATION RESEARCH. Volume 14, Number 1, 2011. ª Mary Ann Liebert, Inc. DOI: 10.1089/rej.2010.1085.
3. Caterini R, Trials to extend MRC5 cell life expectancy induced by a telomerase activator agent. Report in University of Lyon, VALBEX Laboratory, 06/2012.