The 21-day postnatal rat ventricular cardiac muscle cell in culture as an experimental model to study adult cardiomyocyte gene expression

M. L. Lam, Manuela Bartoli, W. C. Claycomb

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

The purpose of this study was to develop and characterize a cardiomyocyte culture system for use as an experimental model to study the mechanism(s) by which cardiac muscle cells permanently exit the cell cycle during early neonatal life. Ventricular cardiomyocytes, isolated by retrograde perfusion of hearts from 21-day-old and adult rats, were compared through 10 days of culture. Expression patterns of genes encoding developmentally programmed proteins were determined to be similar between cardiomyocytes cultured from 21-day-old and adult rats, using the reverse transcription polymerase chain reaction. A lacZ-expressing reporter gene was used to test the efficiency of gene delivery in cultured cardiomyocytes. Transfections using cationic liposomes yielded 24 ± 7, 25 ± 7 and 10 ± 1% cardiomyocytes positive for β-galactosidase activity in cultured 1-day, 21-day and adult cardiomyocytes, respectively. Direct needle microinjection resulted in 48 ± 7, 35 ± 6 and 37 ± 5% cardiomyocytes positive for enzymatic activity in 1-day, 21-day and adult cardiomyocytes, respectively. Cell cycle-specific cDNA arrays were used to analyze the expression pattern of cell cycle-related genes in 12-O-tetradecanoyl-phorbol-13-acetate (TPA)- and non-TPA-treated cultured 21-day cardiomyocytes. Based on the similarity of cultured 21-day to adult ventricular cardiomyocytes and their high transfection efficiencies, we propose the use of cultured cardiomyocytes from 21-day-old rat ventricles as an experimental model system for the study of adult cardiomyocyte gene expression and cell cycle machinery.

Original languageEnglish (US)
Pages (from-to)51-62
Number of pages12
JournalMolecular and Cellular Biochemistry
Volume229
Issue number1-2
DOIs
StatePublished - Mar 12 2002

Fingerprint

Cell culture
Cardiac Myocytes
Gene expression
Muscle
Rats
Theoretical Models
Cell Culture Techniques
Cells
Gene Expression
Genes
Galactosidases
Gene encoding
Polymerase chain reaction
Tetradecanoylphorbol Acetate
Transcription
Liposomes
Cell Cycle
Needles
Machinery
Acetates

Keywords

  • Adult ventricular cardiomyocytes
  • Cell cycle arrays
  • Gene expression
  • Microinjection
  • Transfection

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

The 21-day postnatal rat ventricular cardiac muscle cell in culture as an experimental model to study adult cardiomyocyte gene expression. / Lam, M. L.; Bartoli, Manuela; Claycomb, W. C.

In: Molecular and Cellular Biochemistry, Vol. 229, No. 1-2, 12.03.2002, p. 51-62.

Research output: Contribution to journalArticle

@article{d65acfdb487a4e1d9994f583abbe4c7a,
title = "The 21-day postnatal rat ventricular cardiac muscle cell in culture as an experimental model to study adult cardiomyocyte gene expression",
abstract = "The purpose of this study was to develop and characterize a cardiomyocyte culture system for use as an experimental model to study the mechanism(s) by which cardiac muscle cells permanently exit the cell cycle during early neonatal life. Ventricular cardiomyocytes, isolated by retrograde perfusion of hearts from 21-day-old and adult rats, were compared through 10 days of culture. Expression patterns of genes encoding developmentally programmed proteins were determined to be similar between cardiomyocytes cultured from 21-day-old and adult rats, using the reverse transcription polymerase chain reaction. A lacZ-expressing reporter gene was used to test the efficiency of gene delivery in cultured cardiomyocytes. Transfections using cationic liposomes yielded 24 ± 7, 25 ± 7 and 10 ± 1{\%} cardiomyocytes positive for β-galactosidase activity in cultured 1-day, 21-day and adult cardiomyocytes, respectively. Direct needle microinjection resulted in 48 ± 7, 35 ± 6 and 37 ± 5{\%} cardiomyocytes positive for enzymatic activity in 1-day, 21-day and adult cardiomyocytes, respectively. Cell cycle-specific cDNA arrays were used to analyze the expression pattern of cell cycle-related genes in 12-O-tetradecanoyl-phorbol-13-acetate (TPA)- and non-TPA-treated cultured 21-day cardiomyocytes. Based on the similarity of cultured 21-day to adult ventricular cardiomyocytes and their high transfection efficiencies, we propose the use of cultured cardiomyocytes from 21-day-old rat ventricles as an experimental model system for the study of adult cardiomyocyte gene expression and cell cycle machinery.",
keywords = "Adult ventricular cardiomyocytes, Cell cycle arrays, Gene expression, Microinjection, Transfection",
author = "Lam, {M. L.} and Manuela Bartoli and Claycomb, {W. C.}",
year = "2002",
month = "3",
day = "12",
doi = "10.1023/A:1017999216277",
language = "English (US)",
volume = "229",
pages = "51--62",
journal = "Molecular and Cellular Biochemistry",
issn = "0300-8177",
publisher = "Springer Netherlands",
number = "1-2",

}

TY - JOUR

T1 - The 21-day postnatal rat ventricular cardiac muscle cell in culture as an experimental model to study adult cardiomyocyte gene expression

AU - Lam, M. L.

AU - Bartoli, Manuela

AU - Claycomb, W. C.

PY - 2002/3/12

Y1 - 2002/3/12

N2 - The purpose of this study was to develop and characterize a cardiomyocyte culture system for use as an experimental model to study the mechanism(s) by which cardiac muscle cells permanently exit the cell cycle during early neonatal life. Ventricular cardiomyocytes, isolated by retrograde perfusion of hearts from 21-day-old and adult rats, were compared through 10 days of culture. Expression patterns of genes encoding developmentally programmed proteins were determined to be similar between cardiomyocytes cultured from 21-day-old and adult rats, using the reverse transcription polymerase chain reaction. A lacZ-expressing reporter gene was used to test the efficiency of gene delivery in cultured cardiomyocytes. Transfections using cationic liposomes yielded 24 ± 7, 25 ± 7 and 10 ± 1% cardiomyocytes positive for β-galactosidase activity in cultured 1-day, 21-day and adult cardiomyocytes, respectively. Direct needle microinjection resulted in 48 ± 7, 35 ± 6 and 37 ± 5% cardiomyocytes positive for enzymatic activity in 1-day, 21-day and adult cardiomyocytes, respectively. Cell cycle-specific cDNA arrays were used to analyze the expression pattern of cell cycle-related genes in 12-O-tetradecanoyl-phorbol-13-acetate (TPA)- and non-TPA-treated cultured 21-day cardiomyocytes. Based on the similarity of cultured 21-day to adult ventricular cardiomyocytes and their high transfection efficiencies, we propose the use of cultured cardiomyocytes from 21-day-old rat ventricles as an experimental model system for the study of adult cardiomyocyte gene expression and cell cycle machinery.

AB - The purpose of this study was to develop and characterize a cardiomyocyte culture system for use as an experimental model to study the mechanism(s) by which cardiac muscle cells permanently exit the cell cycle during early neonatal life. Ventricular cardiomyocytes, isolated by retrograde perfusion of hearts from 21-day-old and adult rats, were compared through 10 days of culture. Expression patterns of genes encoding developmentally programmed proteins were determined to be similar between cardiomyocytes cultured from 21-day-old and adult rats, using the reverse transcription polymerase chain reaction. A lacZ-expressing reporter gene was used to test the efficiency of gene delivery in cultured cardiomyocytes. Transfections using cationic liposomes yielded 24 ± 7, 25 ± 7 and 10 ± 1% cardiomyocytes positive for β-galactosidase activity in cultured 1-day, 21-day and adult cardiomyocytes, respectively. Direct needle microinjection resulted in 48 ± 7, 35 ± 6 and 37 ± 5% cardiomyocytes positive for enzymatic activity in 1-day, 21-day and adult cardiomyocytes, respectively. Cell cycle-specific cDNA arrays were used to analyze the expression pattern of cell cycle-related genes in 12-O-tetradecanoyl-phorbol-13-acetate (TPA)- and non-TPA-treated cultured 21-day cardiomyocytes. Based on the similarity of cultured 21-day to adult ventricular cardiomyocytes and their high transfection efficiencies, we propose the use of cultured cardiomyocytes from 21-day-old rat ventricles as an experimental model system for the study of adult cardiomyocyte gene expression and cell cycle machinery.

KW - Adult ventricular cardiomyocytes

KW - Cell cycle arrays

KW - Gene expression

KW - Microinjection

KW - Transfection

UR - http://www.scopus.com/inward/record.url?scp=0036191278&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0036191278&partnerID=8YFLogxK

U2 - 10.1023/A:1017999216277

DO - 10.1023/A:1017999216277

M3 - Article

VL - 229

SP - 51

EP - 62

JO - Molecular and Cellular Biochemistry

JF - Molecular and Cellular Biochemistry

SN - 0300-8177

IS - 1-2

ER -