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. 2016 May 24;17(6):800. doi: 10.3390/ijms17060800

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© 2016 by the authors; licensee MDPI, Basel, Switzerland.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC-BY) license (http://creativecommons.org/licenses/by/4.0/).

PMC Copyright notice

Sca-1+/CD31− CSCs^hTERT CM protects HL-1 cardiomyocytes from hypoxic injury partly via MCP-1-dependent mechanism. Sca-1+/CD31− CSCs^hTERT were cultured in six-well plates at a density of 5 × 10⁴ cells/well and transfected with 50 nM of MCP-1 siRNA duplexes or NC siRNA duplexes using Lipofectamine RNAiMAX. After a 48 h transfection, MCP-1 mRNA expression was assessed by real-time PCR (A); The average value of MCP-1 mRNA was normalized to that of GAPDH for each sample. The data represent the mean ± SD from three experiments. Sca-1+/CD31− CSCs^hTERT CM (B) after transfection of NC siRNA or MCP-1 siRNA were subjected to densitometry and are presented as fold changes for MCP-1 (indicated by number 6) and IL-6 (indicated by number 15) (C), taking MCP-1 and IL-6 levels in NC siRNA-transfected Sca-1+/CD31− CSCs^hTERT as a one-fold value, each in triplicate, * p < 0.05; ** p < 0.01 vs. NC siRNA; (D) representative flow cytometric analysis showing apoptotic effects on HL-1 cardiomyocytes treated with or without 150 μM CoCl₂ for 24 h in the absence or presence of Sca-1+/CD31− CSCs^hTERT CM transfected with NC siRNA or MCP-1 siRNA. Bar graph of three independent experiments showing percentages of apoptotic HL-1 cardiomyocytes (E) treated with or without 150 μM CoCl₂ for 24 h in the absence or presence of Sca-1+/CD31− CSCs^hTERT CM transfected with NC siRNA or MCP-1 siRNA (** p < 0.01); and (F) representative flow cytometric analysis showing apoptotic effects on HL-1 cardiomyocytes treated with or without 200 μM H₂O₂ for 24 h in the absence or presence of Sca-1+/CD31− CSCs^hTERT CM transfected with NC siRNA or MCP-1 siRNA; Bar graph of three independent experiments showing percentages of apoptotic HL-1 cardiomyocytes (G) treated with or without 200 μM H₂O₂ for 24 h in the absence or presence of Sca-1+/CD31− CSCs^hTERT CM transfected with NC siRNA or MCP-1 siRNA (** p < 0.01).

Sca-1+/CD31− CSCs^hTERT CM protects HL-1 cardiomyocytes from hypoxic injury partly via MCP-1-dependent mechanism. Sca-1+/CD31− CSCs^hTERT were cultured in six-well plates at a density of 5 × 10⁴ cells/well and transfected with 50 nM of MCP-1 siRNA duplexes or NC siRNA duplexes using Lipofectamine RNAiMAX. After a 48 h transfection, MCP-1 mRNA expression was assessed by real-time PCR (A); The average value of MCP-1 mRNA was normalized to that of GAPDH for each sample. The data represent the mean ± SD from three experiments. Sca-1+/CD31− CSCs^hTERT CM (B) after transfection of NC siRNA or MCP-1 siRNA were subjected to densitometry and are presented as fold changes for MCP-1 (indicated by number 6) and IL-6 (indicated by number 15) (C), taking MCP-1 and IL-6 levels in NC siRNA-transfected Sca-1+/CD31− CSCs^hTERT as a one-fold value, each in triplicate, * p < 0.05; ** p < 0.01 vs. NC siRNA; (D) representative flow cytometric analysis showing apoptotic effects on HL-1 cardiomyocytes treated with or without 150 μM CoCl₂ for 24 h in the absence or presence of Sca-1+/CD31− CSCs^hTERT CM transfected with NC siRNA or MCP-1 siRNA. Bar graph of three independent experiments showing percentages of apoptotic HL-1 cardiomyocytes (E) treated with or without 150 μM CoCl₂ for 24 h in the absence or presence of Sca-1+/CD31− CSCs^hTERT CM transfected with NC siRNA or MCP-1 siRNA (** p < 0.01); and (F) representative flow cytometric analysis showing apoptotic effects on HL-1 cardiomyocytes treated with or without 200 μM H₂O₂ for 24 h in the absence or presence of Sca-1+/CD31− CSCs^hTERT CM transfected with NC siRNA or MCP-1 siRNA; Bar graph of three independent experiments showing percentages of apoptotic HL-1 cardiomyocytes (G) treated with or without 200 μM H₂O₂ for 24 h in the absence or presence of Sca-1+/CD31− CSCs^hTERT CM transfected with NC siRNA or MCP-1 siRNA (** p < 0.01).