在工程化心脏组织模型中，pH缓冲可减少缺氧性细胞死亡。
Hypoxic cell death is reduced by pH buffering in a model of engineered heart tissue.
作者信息
Brown David A, MacLellan William R, Dunn James C Y, Wu Benjamin M, Beygui Ramin E
机构信息
Department of Bioengineering, University of California, Los Angeles, CA 90095, USA.
出版信息
Artif Cells Blood Substit Immobil Biotechnol. 2008;36(2):94-113. doi: 10.1080/10731190801932090.
INTRODUCTION
In this report, we tested the ability of HEPES-buffered culture medium to reduce acidotic cell death in hypoxic monolayer cell cultures and in a diffusion-limited model of engineered heart tissue (EHT).
METHODS
Neonatal rat cardiomyocytes were either plated (monolayers) or suspended in a hydrogel disc containing HEPES.
RESULTS
Monolayers cultured in 0% oxygen exhibited a pH drop to 5.46 +/- 0.27 after 4 days with no HEPES, or 7.11 +/- 0.09 with 50 mM HEPES. The lowest observed pH in EHTs was estimated as 6.2 with no HEPES and 7.1 with 50 mM HEPES, which were endpoints of noticeably different pH gradients across the EHTs. Addition of HEPES to hypoxic monolayers corresponded with fewer propidium iodide-positive cells and TUNEL-positive cells; addition of HEPES to EHTs resulted in greater calcein staining and less LDH release.
CONCLUSION
Effective pH buffering reduces cell death by attenuating the acidosis that accompanies anaerobic metabolism.
引言
在本报告中，我们测试了HEPES缓冲培养基在缺氧单层细胞培养物和工程化心脏组织（EHT）的扩散限制模型中减少酸中毒细胞死亡的能力。
方法
将新生大鼠心肌细胞接种（单层培养）或悬浮于含有HEPES的水凝胶盘中。
结果
在无氧条件下培养4天后，未添加HEPES的单层细胞pH降至5.46±0.27，而添加50 mM HEPES的单层细胞pH为7.11±0.09。在EHT中，未添加HEPES时观察到的最低pH估计为6.2，添加50 mM HEPES时为7.1，这是EHT中明显不同pH梯度的终点。向缺氧单层细胞中添加HEPES可使碘化丙啶阳性细胞和TUNEL阳性细胞减少；向EHT中添加HEPES可使钙黄绿素染色增强，乳酸脱氢酶释放减少。
结论
有效的pH缓冲通过减轻无氧代谢伴随的酸中毒来减少细胞死亡。
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