National Academy of Sciences of Ukraine Bogomoletz Institute of Physiology The influence of hydrogen sulfide donor and oxygen partial pressure on the mesenchymal stem cells proliferation Lidiia Plotnikova

What Are Stem Cells? STEM CELLS- cells which can make exact copies of itself indefinitely, can differentiate, and produce specialized cells for various tissues of body.

Factors affecting cell proliferation in vitro The composition of the gas environment (О2, СО2, Н2S etc.) The composition of the culture medium (amino acids, vitamins, glucose, growth factors etc.) Proliferation Physico-chemical properties of the medium (рН, t°C, osmolarity etc.) The density monolayer, intercellular interaction 3 / 8

The aim and scheme of the experiment The aim of our work was to investigate the effect of hydrogen sulfide donor and low partial oxygen pressure (Ро2) on proliferation of human cell line 4BL. Cell line 4BL − multipotent mesenchymal stromal cells (MSC) derived from the blood of healthy donor and translated in conditions of standard monolayer culture in Institute of Molecular Biology and Genetics National Academy of Sciences of Ukraine. Impact of hydrogen sulfide donor (NaHS) in concentrations: 10-8 , 10-9 , 10-10, 10-12, 10-13 , 10-15 mol / l. Ро2 – Control 159 mmHg (21% O2) – 38 mmHg (5% O2) – 76 mmHg (10% O2) Plated 50 thousand сells Mode of incubation: I – 24 h II – 8 h in day (x3) counted the number of cells after 72 and 96 hours d=35 mm

Cell line 4BL proliferation rights under the addition of hydrogen sulfide donor on the third day of cultivation N•103, cells/ml СNaHS, mol/l 200 мкм 200 мкм 200 мкм 200 мкм Painting with Romanovsky-Himzoyu. ×100. *р<0,05 – statistical probability compared with control 5 / 8

Proliferation of human MSC at a lessened partial oxygen pressure Mode of incubation: I - 24 h II - 8 h in day (×3 ) 72 h 96 h *р<0,05 – statistical probability compared with control

The cells line 4BL on the 4th day of cultivation in control and at 24-hour Po2 of 76 mm Hg 100 мкм 200 мкм Po2 of 76 mm Hg 100 мкм 200 мкм Painting with Romanovsky-Himzoyu. Microscope increase ×100 and ×400 7 / 8

Conclusion The hydrogen sulfide donor in concentrations 10-8-10-10 mol / l has cytotoxic effect on cell culture and in concentrations 10-13-10-15 mol / l - inhibits proliferation. Cultivation MSC line 4BL in a gaseous medium with a low oxygen concentration (5 % and 10 % O2) for 8 hours per day (3 times) significantly increases the cell proliferation by 27 % and 45 % respectively.