The effects of cysplatin on human adipose tissue derived mesenchymal stromal cells under different oxygen levels

Abstract

To evaluate the damaging effects of cisplatin on MMSCs from adipose tissue in a phase of active proliferation and the state of the monolayer, this was exposed at standard (20%) and reduced to 1% and 5% level of oxygen. The effect of cisplatin was detected on monolayer cultures in the active growth phase after 2 passages. Profile surface markers of MMSC was determined by flow cytometry. MMSCs viability after incubation with cisplatin was detected by the number of apoptotic and necrotic cells using ANNEXIN V-FITC--PI Kit (Immunotech, France). Standard culture conditions (~20% O₂) were created in a CO₂ incubator (Sanyo, Japan), 5% O₂--using multigas incubator (Sanyo, Japan), 1% O₂--using an airtight chamber (Stemcell Technologies, USA). Incubation of MMSC monolayer with cisplatin at a concentration of 10 ug/ml for 72 hours leads to death of half of the cells in the culture. Cisplatin increased thefracture of PI⁺-cell, and PI⁺/Ann⁺-cells under all culture conditions. The short-term exposure with cisplatin (24 and 48 hours) did not cause the damaging effect. Effects of cisplatin on the MMSC in the growth phase for 48 hours led to accumulation of Ann⁺-cells and PI⁺/Ann⁺-cells under all culture conditions. However, minimal noci-influence of cisplatin was observed in a culture under hypoxic conditions (1% O₂). These data suggest that MMSC monolayer dies primarily through necrosis, whereas MMSCs in the growth phase in response to cisplatin treatment dies by apoptosis, regardless the oxygen tension.