TY - JOUR
T1 - An antegrade perfusion method for cardiomyocyte isolation from mice
AU - Omatsu-Kanbe, Mariko
AU - Fukunaga, Ryo
AU - Mi, Xinya
AU - Matsuura, Hiroshi
N1 - Funding Information:
The authors thank T. Yamamoto and Y. Mori for their assistance in the morphologic experiments. This work was supported by a Grant-in-Aid for Scientific Research (C) from the Japan Society for the Promotion of Science (18K06871 to M.O.K. and 17K08536 to H.M.).
Publisher Copyright:
© 2021, Journal of Visualized Experiments. All rights reserved.
PY - 2021
Y1 - 2021
N2 - In basic research using mouse heart, isolating viable individual cardiomyocytes is a crucial technical step to overcome. Traditionally, isolating cardiomyocytes from rabbits, guinea pigs or rats has been performed via retrograde perfusion of the heart with enzymes using a Langendorff apparatus. However, a high degree of skill is required when this method is used with a small mouse heart. An antegrade perfusion method that does not use a Langendorff apparatus was recently reported for the isolation of mouse cardiomyocytes. We herein report a complete protocol for the improved antegrade perfusion of the excised heart to isolate individual heart cells from adult mice (8-108 weeks old). Antegrade perfusion is performed by injecting perfusate near the apex of the left ventricle of the excised heart, the aorta of which was clamped, using an infusion pump. All procedures are carried out on a pre-warmed heater mat under a microscope, which allows for the injection and perfusion processes to be monitored. The results suggest that ventricular and atrial myocytes, and fibroblasts can be well isolated from a single adult mouse simultaneously.
AB - In basic research using mouse heart, isolating viable individual cardiomyocytes is a crucial technical step to overcome. Traditionally, isolating cardiomyocytes from rabbits, guinea pigs or rats has been performed via retrograde perfusion of the heart with enzymes using a Langendorff apparatus. However, a high degree of skill is required when this method is used with a small mouse heart. An antegrade perfusion method that does not use a Langendorff apparatus was recently reported for the isolation of mouse cardiomyocytes. We herein report a complete protocol for the improved antegrade perfusion of the excised heart to isolate individual heart cells from adult mice (8-108 weeks old). Antegrade perfusion is performed by injecting perfusate near the apex of the left ventricle of the excised heart, the aorta of which was clamped, using an infusion pump. All procedures are carried out on a pre-warmed heater mat under a microscope, which allows for the injection and perfusion processes to be monitored. The results suggest that ventricular and atrial myocytes, and fibroblasts can be well isolated from a single adult mouse simultaneously.
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U2 - 10.3791/61866
DO - 10.3791/61866
M3 - Article
C2 - 34096909
AN - SCOPUS:85107675596
SN - 1940-087X
VL - 2021
JO - Journal of Visualized Experiments
JF - Journal of Visualized Experiments
IS - 171
M1 - e61866
ER -