A Transgenic tdTomato Rat for Cell Migration and Tissue Engineering Applications

Tissue Engineering. Part C, Methods
Brian C SyverudLisa Marie Larkin

Abstract

The growing deficit in suitable tissues for patients awaiting organ transplants demonstrates the clinical need for engineered tissues as alternative graft sources. Demonstrating safety and efficacy by tracking the migration and fate of implanted cells is a key consideration required for approval of promising engineered tissues. Cells from transgenic animals that express green fluorescent protein (GFP) are commonly used for this purpose. However, GFP can create difficulties in practice due to high levels of green autofluorescence in many musculoskeletal tissues. Tandem-dimer tomato (tdTomato) is a stable, robust red fluorescent protein that is nearly threefold brighter than GFP. Our objective was to create a line of transgenic rats that ubiquitously express tdTomato in all cells, driven by the human ubiquitin C promoter. We sought to determine the rats' utility in tissue engineering applications by fabricating engineered skeletal muscle units (SMUs) from isolated muscle-derived tdTomato cells. These tdTomato SMUs were implanted into a volumetric muscle loss (VML) defect of the tibialis anterior muscle in a rat ubiquitously expressing GFP. We also evaluated a novel method for modularly combining individual SMUs to create a larger...Continue Reading

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Citations

Apr 8, 2020·Regenerative Engineering and Translational Medicine·Emmanuel E Vega-SotoLisa M Larkin
Jun 21, 2019·Regenerative Engineering and Translational Medicine·Brittany L RodriguezLisa M Larkin
Feb 6, 2020·Tissue Engineering. Part a·Stoyna S NovakovaLisa M Larkin
Nov 19, 2020·Tissue Engineering. Part a·Olga M WroblewskiLisa M Larkin

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Methods Mentioned

BETA
transfection
FRET
transgenic
genetic modification
biopsies
PCR
Genotyping
dissection

Software Mentioned

ImageJ
EVOS
Living Image
GraphPad Prism

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