An improved method for primary culture of neonatal mouse cardiomyocytes
10.3969/j.issn.1671-7856.2016.04.011
- VernacularTitle:改良的乳小鼠心肌细胞原代培养方法
- Author:
Dan WU
;
Jian FENG
;
Xiangang MO
;
Yingcai LIU
- Publication Type:Journal Article
- Keywords:
Neonatal mouse;
Cardiomyocytes;
Primary cell culture
- From:
Chinese Journal of Comparative Medicine
2016;26(4):62-67
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a stable and fast method for primary culture of mouse cardiomyocytes. Methods Dishes were coated with polylysine firstly.A two-step approach was used to isolate and digest mouse cardiomyocytes cells (0.25%trypsin in 4°C overnight and 0.5 mg/mL to 1.0 mg/mL collagenase +5 mg/mL albumin collagen digestion liquid in 37°C for short-time digestion), then the cardiomyocytes were purified through differential adhesion for 70 min and 5-bromodeoxyuridine ( BrdU) .The cell morphology was observed under an inverted microscope. The survival rate of cardiacmyocytes was detected by trypan-blue staining and their purity was identified by α-actinin immunofluorescence staining.Results The cardiomyocytes were in good shape and pulsed spontaneously.The survival rate of the cardiomyocytes reached 98%and the purity was 95%.Conclusions This method described in this study is an ideal method for primary culture of mouse cardiomyocytes with a high survival rate and high purity.
