Production of Spontaneously Beating Neonatal Rat Heart Tissue for Calcium and Contractile Studies View Full Text


Ontology type: schema:Chapter     


Chapter Info

DATE

2013

AUTHORS

Fnu Gerilechaogetu , Hao Feng , Honey B. Golden , Damir Nizamutdinov , Donald M. Foster , Shannon Glaser , David E. Dostal

ABSTRACT

Neonatal rat ventricular myocytes (NRVM) and fibroblasts (FB) serve as in vitro models for studying fundamental mechanisms underlying cardiac pathologies, as well as identifying potential therapeutic targets. Typically, these cell types are separated using Percoll density gradient procedures. Cells located between the Percoll bands (interband cells [IBCs]), which contain less mature NRVM and a variety of non-myocytes, including coronary vascular smooth muscle cells and endothelial cells (ECs), are routinely discarded. However, we have demonstrated that IBCs readily attach to extracellular matrix-coated coverslips, plastic culture dishes, and deformable membranes to form a 2-dimensional cardiac tissue layer which quickly develops spontaneous contraction within 24 h, providing a robust coculture model for the study of cell-to-cell signaling and contractile studies. Below, we describe methods that provide good cell yield and viability of IBCs during isolation of NRVM and FB obtained from 0- to 3-day-old neonatal rat pups. Basic characterization of IBCs and methods for use in intracellular calcium and contractile experiments are also presented. This method maximizes the use of cells obtained from neonatal rat hearts. More... »

PAGES

45-56

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/978-1-62703-604-7_4

DOI

http://dx.doi.org/10.1007/978-1-62703-604-7_4

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1053529377

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/23955732


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