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Quantitative assessment of cardiomyocyte mechanobiology through high-throughput cantilever-based functional well plate systems.
Kim, Jongyun; Shanmugasundaram, Arunkumar; Kim, Dong-Su; Jeong, Yun-Jin; Kanade, Pooja P; Kim, Eung-Sam; Lee, Bong-Kee; Lee, Dong-Weon.
Affiliation
  • Kim J; MEMS and Nanotechnology Laboratory, School of Mechanical Engineering, Chonnam National University, Gwangju 61186, Republic of Korea. mems@jnu.ac.kr.
  • Shanmugasundaram A; Advanced Medical Device Research Center for Cardiovascular Disease, Chonnam National University, Gwangju 61186, Republic of Korea.
  • Kim DS; MEMS and Nanotechnology Laboratory, School of Mechanical Engineering, Chonnam National University, Gwangju 61186, Republic of Korea. mems@jnu.ac.kr.
  • Jeong YJ; Advanced Medical Device Research Center for Cardiovascular Disease, Chonnam National University, Gwangju 61186, Republic of Korea.
  • Kanade PP; MEMS and Nanotechnology Laboratory, School of Mechanical Engineering, Chonnam National University, Gwangju 61186, Republic of Korea. mems@jnu.ac.kr.
  • Kim ES; Advanced Medical Device Research Center for Cardiovascular Disease, Chonnam National University, Gwangju 61186, Republic of Korea.
  • Lee BK; Green Energy & Nano Technology R&D Group, Korea Institute of Industrial Technology (KITECH), Gwangju, 61012, Republic of Korea.
  • Lee DW; MEMS and Nanotechnology Laboratory, School of Mechanical Engineering, Chonnam National University, Gwangju 61186, Republic of Korea. mems@jnu.ac.kr.
Analyst ; 148(20): 5133-5143, 2023 Oct 05.
Article in En | MEDLINE | ID: mdl-37695027
ABSTRACT
Proper regulation of the in vitro cell culture environment is essential for disease modelling and drug toxicity screening. The main limitation of well plates used for cell culture is that they cannot accurately maintain energy sources and compounds needed during cell growth. Herein, to understand the importance of perfusion in cardiomyocyte culture, changes in contractile force and heart rate during cardiomyocyte growth are systematically investigated, and the results are compared with those of a perfusion-free system. The proposed perfusion system consists of a Peltier refrigerator, a peristaltic pump, and a functional well plate. A functional well plate with 12 wells is made through injection moulding, with two tubes integrated in the cover for each well to continuously circulate the culture medium. The contractile force of cardiomyocytes growing on the cantilever surface is analysed through changes in cantilever displacement. The maturation of cardiomyocytes is evaluated through fluorescence staining and western blot; cardiomyocytes cultured in the perfusion system show greater maturity than those cultured in a manually replaced culture medium. The pH of the culture medium manually replaced at intervals of 3 days decreases to 6.8, resulting in an abnormal heartbeat, while cardiomyocytes cultured in the perfusion system maintained at pH 7.4 show improved contractility and a uniform heart rate. Two well-known ion channel blockers, verapamil and quinidine, are used to measure changes in the contractile force of cardiomyocytes from the two systems. Cardiomyocytes in the perfusion system show greater stability during drug toxicity screening, proving that the perfusion system provides a better environment for cell growth.
Subject(s)

Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Myocytes, Cardiac / Drug-Related Side Effects and Adverse Reactions Limits: Humans Language: En Journal: Analyst Year: 2023 Type: Article

Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Myocytes, Cardiac / Drug-Related Side Effects and Adverse Reactions Limits: Humans Language: En Journal: Analyst Year: 2023 Type: Article