4.3 Article

Improvements in the primary culture of neonate rat myocardial cells by study of the mechanism of endoplasmic reticulum stress

期刊

CELL STRESS & CHAPERONES
卷 18, 期 3, 页码 367-375

出版社

SPRINGER
DOI: 10.1007/s12192-012-0390-y

关键词

Endoplasmic reticulum stress; Primary culture of neonate rat myocardial cells; Culture technique

资金

  1. National Science Foundation for Young Scholars of China [30900612]
  2. Medical Science Research Foundation of Zhejiang Province [2009B127]
  3. Zhejiang Provincial Natural Science Foundation, People's Republic of China [Y2090393]

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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.

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