THE EFFECT OF MRET ACTIVATED MEDIUM ON CELL MORPHOLOGY

THE EFFECT OF MRET ACTIVATED MEDIUM ON CELL MORPHOLOGY

The in vitro investigations on normal PMBC cells (peripheral blood mononuclear cell) and on HeLa cancer cells (cell line ATCC # CCL-2 cervical adenocarcinoma) and were conducted under the supervision of Patrick Pezzoli, Ph.D. at AltheaDx Technologies, San Diego, USA. The experiments analyzed: cells lysed at 0 hour, cells cultured for 24 hours in untreated medium and cells cultured for 24 hours in medium treated with MRET activator for 30 minutes. DNA samples from each batch were processed and the resultant data was analyzed using Affymetrix Genotyping Console 3.0 to obtain genotype calls and copy number calls. Cell counts and % viability were obtained using the Trypan Blue exclusion technique.

The experimental data revealed no difference between the zero hour (control), MRET treated and untreated samples in term of genotypes and copy number calls. Thus, MRET activation of water based medium did not induce any changes in cells on genetic level.

The studies showed that in MRET activated medium the viability of normal cells (PBMC) was higher (Table 1), and the viability of cancer cells (HeLa) was lower (Table 2) compared to their viability in untreated medium.

Sample  Cell Count   % Viability  Viable cells
0 hour   3.27x106 91.5 2.99x106
Untreated   8.70x105 83.4 0.73x106
Treated 7.73x105 88.8 0.69x106

Table 1: PBMC cell counts and % viability

Sample  Cell Count   % Viability  Viable cells
0 hour   3x106 92  3.27x106
Untreated   7x106   97  6.79x106
Treated 5x106 92  4.60x106

Table 2: HeLa cell counts and % viability


For normal cells (PBMC) the changes in cell counts were similar for untreated and MRET treated medium (Fig 1). Thus, MRET treatment did not affect the growth of normal cells.

For cancer cells (HeLa) the experimental data revealed significant inhibition of cancer cells growth in MRET treated medium. The growth of viable cancer cells was inhibited by 54% in MRET treated medium compared to untreated medium (Fig 2).

The results of AltheaDx research on HeLa cancer cells in vitro support the results obtained earlier in the investigation regarding the effects of MRET water on tumor resistance in animal model. The study on 500 mice was conducted under the supervision of Professor V. Vysotskii, S. Olishevsky, Ph.D. and Y. Yanish, Ph.D. at Kyiv Institute of Experimental Pathology, Oncology and Radiobiology of Ukrainian Academy of Science. It showed substantial inhibition of growth of viable tumor cells following the consumption of MRET water. In the course of this investigation the groups of mice in “preventive regime” ingested MRET water for 2 weeks prior to the inoculation of Ehrlich carcinoma cancer cells and for 3 weeks after the inoculation. The groups of mice in “therapeutic regime” ingested MRET water only during 3 weeks after the inoculation of Ehrlich carcinoma cancer cells. Following the consumption of MRET water activated for 30 minutes (the optimal time of activation) the growth of viable tumor cells was inhibited by 76% in “preventive regime” and by 55% in “therapeutic regime.”

As a conclusion it is possible to say that the studies conducted at AltheaDx Technology confirmed that MRET activated water did not affect the cells on genetic level; it affected the morphology of normal cells in positive way increasing their viability and promoted significant inhibition of growth of cancer cells.