Temnov Andrey, Rogov Konstantin, Zhalimov Vitaliy, Igor Popov, Pekov Stanislav, Bader Augustinus, Sklifas Alla, Giri Shibashish
Department of Biology, Institute of Cell Biophysics of the Russian Academy of Sciences, Pushchino, Russia.
Department of Biotechnology, Moscow Institute of Physics and Technology, Dolgoprudny, Russia.
Hepat Med. 2019 Jul 19;11:89-96. doi: 10.2147/HMER.S196354. eCollection 2019.
In our studies, it was shown that the effectiveness of the conditioned medium obtained by cultivating mesenchymal stem cells depends on the microenvironment conditions used to cultivate the cells. It was demonstrated that the conditioned medium obtained by culturing cells with low oxygen content (10%) has a much more pronounced protective effect. Protein compositions obtained from MSCs cultured under hypoxic (10% O hc-MSC) and normal (21% O nc-MSC) conditions were used to treat acute liver failure (ALF) induced in mice by acetaminophen injection. Thus, we obtained fractions normalized by volume, which predominantly contained proteins with masses > 50, 50-30, 30-10, and 10-3 kDa. The data from biochemical studies have shown that only fractions from 10 to 30 kDa (hcMSC and ncMSC) significantly reduced the level of liver enzymes in the beginning of the acute period after acetaminophen administration. Mass spectrometry analysis of the proteins contained in the isolated fractions showed a sharp increase in the protein levels in the 10-30 kDa hcMSC fraction as compared with that in 10-30 kDa ncMSCs. The composition obtained from MSCs cultured at lower O level (fraction 10-30 kDa hcMSC) was shown to be more potent than the composition prepared from normoxic cells. The results have shown that a composition obtained by culturing the cells under a reduced content of O (10%), significantly improves the biochemical parameters, and histological arrester reduces the degree of inflammation and stimulates regenerative processes in liver, compared to both the control group and group treated with the composition that was obtained by culturing the cells under normal oxygen content.
在我们的研究中，结果表明通过培养间充质干细胞获得的条件培养基的有效性取决于用于培养细胞的微环境条件。结果表明，在低氧含量（10%）条件下培养细胞获得的条件培养基具有更为显著的保护作用。将在低氧（10% O₂，hc-MSC）和正常（21% O₂，nc-MSC）条件下培养的间充质干细胞获得的蛋白质组合物用于治疗对乙酰氨基酚注射诱导的小鼠急性肝衰竭（ALF）。因此，我们获得了按体积归一化的组分，这些组分主要包含质量大于50、50-30、30-10和10-3 kDa的蛋白质。生化研究数据表明，只有10至30 kDa的组分（hcMSC和ncMSC）在给予对乙酰氨基酚后的急性期开始时显著降低了肝酶水平。对分离组分中所含蛋白质的质谱分析表明，与10-30 kDa的ncMSCs相比，10-30 kDa的hcMSC组分中的蛋白质水平急剧增加。结果表明，从低氧水平培养的间充质干细胞获得的组合物（10-30 kDa hcMSC组分）比从常氧细胞制备的组合物更有效。结果表明，与对照组和用正常氧含量培养细胞获得的组合物处理的组相比，在低氧含量（10%）下培养细胞获得的组合物显著改善了生化参数，组织学检查减少了炎症程度并刺激了肝脏的再生过程。
