title: Improvements in the primary culture of neonate rat myocardial cells by study of the mechanism of endoplasmic reticulum stress.
abstract: 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.
authors: Department of Cardiology, The First Affiliated Hospital, College of Medicine, Zhejiang University, 79 Qingchun Road, Hangzhou, Zhejiang Province, People's Republic of China.
authors: Qiqi
authors: Q
authors: Wang
authors: Jianmei
authors: J
authors: Jin
authors: Zhidong
authors: Z
authors: Guo
authors: Fuxu
authors: F
authors: Chen
authors: Yuangang
authors: Y
authors: Qiu
authors: Jianhua
authors: J
authors: Zhu
authors: Yunpeng
authors: Y
authors: Shang
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D015153:Blotting, Western; D018929:Cell Culture Techniques / Q000379:methods*; D048430:Cell Shape / Q000187:drug effects; D002478:Cells, Cultured; D003470:Culture Media / Q000494:pharmacology; D000091342:Endoplasmic Reticulum Chaperone BiP; D059865:Endoplasmic Reticulum Stress / Q000187:drug effects; D005786:Gene Expression Regulation / Q000187:drug effects; D005947:Glucose / Q000494:pharmacology; D006339:Heart Rate / Q000187:drug effects; D006360:Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D009206:Myocardium / Q000166:cytology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D014415:Tunicamycin / Q000494:pharmacology
chemical_list: D003470:Culture Media; D000091342:Endoplasmic Reticulum Chaperone BiP; D006360:Heat-Shock Proteins; D014415:Tunicamycin; D005947:Glucose