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Fig. 6 | Stem Cell Research & Therapy

Fig. 6

From: Reduced culture temperature attenuates oxidative stress and inflammatory response facilitating expansion and differentiation of adipose-derived stem cells

Fig. 6

Culture of human subcutaneous ASCs at 35 °C attenuates ROS accumulation and apoptosis without affecting cell expansion. Human ASCs were isolated from lipoaspirates, seeded, and immediately cultured either at a normal culture temperature (37 °C) or at a mildly reduced temperature (35 °C). A Comparison of ROS accumulation between ASCs isolated from the same lipoaspirate and grown at either 37 °C or 35 °C was performed by DCFDA staining and its analysis by flow cytometry, at the indicated passages (P). (AI) Representative FACS analyses. (AII) Summary of three independent repeats from three different patients, at P2–4, and one repeat at P5. Data are presented as the mean ± standard deviation. B Cells were counted at each passage and reseeded at equal numbers. Cell counts at each passage are presented (n = 4 independent repeats). c Comparison of the percentage of apoptotic ASCs at each passage of cells grown at 37 °C and 35 °C was performed by PI/Annexin staining, followed by FACS analysis. (CI) A representative FACS analysis. (CII) Summary results of three biological repeats from one patient at P5. Similar differences in the level of apoptosis between 37 and 35 °C ASC cultures were obtained from three independent patients but at different passages. DF RNA was extracted at different passages and the level of expression of the indicated genes was compared between 37 and 35 °C ASCs (n = 2). Data are presented as the mean ± standard deviation. ***p < 0.001; **p < 0.01; *p < 0.05

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