Bioluminescence imaging for assessment of immune responses following implantation of engineered heart tissue (EHT)

J Vis Exp. 2011 Jun 1:(52):2605. doi: 10.3791/2605.

Abstract

Various techniques of cardiac tissue engineering have been pursued in the past decades including scaffolding strategies using either native or bioartificial scaffold materials, entrapment of cardiac myocytes in hydrogels such as fibrin or collagen and stacking of myocyte monolayers. These concepts aim at restoration of compromised cardiac function (e.g. after myocardial infarction) or as experimental models (e.g. predictive toxicology and substance screening or disease modelling). Precise monitoring of cell survival after implantation of engineered heart tissue (EHT) has now become possible using in-vivo bioluminescence imaging (BLI) techniques. Here we describe the generation of fibrin-based EHT from a transgenic rat strain with ubiquitous expression of firefly luciferase (ROSA/luciferase-LEW Tg; ). Implantation is performed into the greater omentum of different rat strains to assess immune responses of the recipient organism following EHT implantation. Comparison of results generated by BLI and the Enzyme Linked Immuno Spot Technique (ELISPOT) confirm the usability of BLI for the assessment of immune responses.

Publication types

  • Research Support, Non-U.S. Gov't
  • Video-Audio Media

MeSH terms

  • Animals
  • Fibrin / chemistry
  • Heart Transplantation / methods*
  • Luciferases, Firefly / biosynthesis
  • Luciferases, Firefly / genetics
  • Luminescent Measurements / methods*
  • Muscle Cells / immunology
  • Muscle Cells / metabolism
  • Muscle Cells / physiology
  • Muscle Cells / transplantation
  • Myocardium / cytology
  • Myocardium / immunology
  • Myocardium / metabolism
  • Rats
  • Rats, Transgenic
  • Tissue Engineering / methods*
  • Tissue Transplantation / methods*

Substances

  • Fibrin
  • Luciferases, Firefly