Isolation, characterization, and differentiation of cardiac stem cells from the adult mouse heart

Santosh K. Yadav, Paras Kumar Mishra

Research output: Contribution to journalArticle

Abstract

Myocardial infarction (MI) is a leading cause of morbidity and mortality around the world. A major goal of regenerative medicine is to replenish the dead myocardium after MI. Although several strategies have been used to regenerate myocardium, stem cell therapy remains a major approach to replenish the dead myocardium of an MI heart. Accumulating evidence suggests the presence of resident cardiac stem cells (CSCs) in the adult heart and their endocrine and/or paracrine effects on cardiac regeneration. However, CSC isolation and their characterization and differentiation toward myocardial cells, especially cardiomyocytes, remains a technical challenge. In the present study, we provided a simple method for the isolation, characterization, and differentiation of CSCs from the adult mouse heart. Here, we describe a density gradient method for the isolation of CSCs, where the heart is digested by a 0.2% collagenase II solution. To characterize the isolated CSCs, we evaluated the expression of CSCs/cardiac markers Sca-1, NKX2-5, and GATA4, and pluripotency/stemness markers OCT4, SOX2, and Nanog. We also determined the proliferation potential of isolated CSCs by culturing them in a Petri dish and assessing the expression of the proliferation marker Ki-67. For evaluating the differentiation potential of CSCs, we selected seven- to ten-days cultured CSCs. We transferred them to a new plate with a cardiomyocyte differentiation medium. They are incubated in a cell culture incubator for 12 days, while the differentiation medium is changed every three days. The differentiated CSCs express cardiomyocyte-specific markers: actinin and troponin I. Thus, CSCs isolated with this protocol have stemness and cardiac markers, and they have a potential for proliferation and differentiation toward cardiomyocyte lineage.

Original languageEnglish (US)
Article numbere58448
JournalJournal of Visualized Experiments
Volume2019
Issue number143
DOIs
StatePublished - Jan 1 2019

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Adult Stem Cells
Stem cells
Stem Cells
Cardiac Myocytes
Myocardium
Myocardial Infarction
Actinin
Incubators
Troponin I
Regenerative Medicine
Cell Separation
Collagenases
Cell- and Tissue-Based Therapy
Gradient methods
Regeneration
Cell culture
Cell Culture Techniques
Morbidity

Keywords

  • Biology
  • GATA4 actinin
  • Issue 143
  • Ki-67
  • NKX2-5
  • Nanog
  • OCT4
  • Sca-1
  • Troponin I

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Cite this

Isolation, characterization, and differentiation of cardiac stem cells from the adult mouse heart. / Yadav, Santosh K.; Mishra, Paras Kumar.

In: Journal of Visualized Experiments, Vol. 2019, No. 143, e58448, 01.01.2019.

Research output: Contribution to journalArticle

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