通过研究内质网应激机制提高新生大鼠心肌细胞原代培养的质量。
Improvements in the primary culture of neonate rat myocardial cells by study of the mechanism of endoplasmic reticulum stress.
机构信息
Department of Cardiology, The First Affiliated Hospital, College of Medicine, Zhejiang University, 79 Qingchun Road, Hangzhou, Zhejiang Province, People's Republic of China.
出版信息
Cell Stress Chaperones. 2013 May;18(3):367-75. doi: 10.1007/s12192-012-0390-y. Epub 2013 Jan 18.
We previously found that endoplasmic reticulum stress (ERS) might be exhibited in the conventional protocol of the primary culture of neonate rat myocardial cells (NRMCs) and that the high glucose concentration (25 mmol/L) in the culture medium might be the cause. Here, we investigated if the high concentration of glucose might influence ERS in myocardial cells during culture. GRP78 expression (ERS marker) was similar in groups with tunicamycin (TM) and without TM in high glucose cultured cells (p > 0.01). Different glucose concentrations elicited different GRP78 expressions according to analyses of protein and RNA levels, which showed ERS in H/H groups. Finally, we found that GRP78 expression was higher in TM groups compared with M/M groups (p < 0.01). The conventional high-glucose culture media during primary culture of NRMCs induced ERS. We propose that medium-glucose culture media should be used and describe an improved protocol for the primary culture of NRMCs.
我们之前发现，内质网应激（ERS）可能在新生大鼠心肌细胞（NRMCs）原代培养的常规方案中表现出来，并且培养基中的高葡萄糖浓度（25mmol/L）可能是其原因。在这里，我们研究了高浓度葡萄糖是否会在培养过程中影响心肌细胞中的 ERS。高葡萄糖培养细胞中，用衣霉素（TM）和不用 TM 的组 GRP78 表达（ERS 标志物）相似（p＞0.01）。根据蛋白和 RNA 水平的分析，不同的葡萄糖浓度会引起不同的 GRP78 表达，表明 H/H 组存在 ERS。最后，我们发现 TM 组的 GRP78 表达高于 M/M 组（p＜0.01）。NRMCs 原代培养中常规高糖培养基会诱导 ERS。我们建议使用中糖培养基，并描述一种改良的 NRMCs 原代培养方案。
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