Monitoring collagen transcription by vascular smooth muscle cells in fibrin-based tissue constructs.
Current methods for measuring collagen content in engineered tissues are incompatible with monitoring of collagen production because they require destruction of the tissue. We have implemented a luciferase-based strategy to monitor collagen production noninvasively. Fibrin-based tissue constructs made using vascular smooth muscle cells stably transfected with a collagen I promoter/luciferase transgene developed with collagen content comparable to control cells, but could be imaged noninvasively to follow collagen transcription during tissue growth in vitro. We showed that these cells reported collagen I production at the transcriptional level in response to the growth factor transforming growth factor-beta1 and fibrinolytic inhibition by epsilon-aminocaproic acid and that these changes were consistent with changes at the mRNA and protein levels. As these cells report collagen changes instantly and without tissue destruction, they will facilitate construct optimization using multiple stimuli to produce functional engineered tissues.
Weinbaum
Justin S
JS
Department of Biomedical Engineering, University of Minnesota , Minneapolis, MN, USA.
Qi
Jie
J
Tranquillo
Robert T
RT
D001483:Base Sequence; D001672:Biocompatible Materials; D003094:Collagen / Q000235:genetics*; D017931:DNA Primers; D005337:Fibrin; D005453:Fluorescence; D006801:Humans; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014158:Transcription, Genetic; D014162:Transfection
D001672:Biocompatible Materials; D017931:DNA Primers; D005337:Fibrin; D003094:Collagen