Bioluminescence-mediated longitudinal monitoring of adipose-derived stem cells in a large mammal ex vivo organ culture.
Recently, ex vivo three-dimensional organ culture systems have emerged to study the physiology and pathophysiology of human organs. These systems also have potential as a translational tool in tissue engineering; however, this potential is limited by our ability to longitudinally monitor the fate and action of cells used in regenerative therapies. Therefore, we investigated luciferase-mediated bioluminescence imaging (BLI) as a non-invasive technique to continuously monitor cellular behavior in ex vivo whole organ culture. Goat adipose-derived stem cells (gADSCs) were transduced with either Firefly luciferase (Fluc) or Gaussia luciferase (Gluc) reporter genes and injected in isolated goat intervertebral discs (IVD). Luciferase activity was monitored by BLI for at least seven days of culture. Additionally, possible confounders specific to avascular organ culture were investigated. Gluc imaging proved to be more suitable compared to Fluc in monitoring gADSCs in goat IVDs. We conclude that BLI is a promising tool to monitor spatial and temporal cellular behavior in ex vivo organ culture. Hence, ex vivo organ culture systems allow pre-screening and pre-validation of novel therapeutic concepts prior to in vivo large animal experimentation. Thereby, organ culture systems can reduce animal use, and improve the speed of innovation by overcoming technological, ethical and financial challenges.
Department of Orthopedic Surgery, VU University Medical Center, MOVE Research Institute Amsterdam, Amsterdam, The Netherlands.
Mirte
M
Peeters
Department of Neurosurgery, VU University Medical Center, Amsterdam, The Netherlands.
Sjoerd
S
van Rijn
Department of Orthopedic Surgery, VU University Medical Center, MOVE Research Institute Amsterdam, Amsterdam, The Netherlands.
Pieter-Paul A
PP
Vergroesen
Department of Orthopedic Surgery, VU University Medical Center, MOVE Research Institute Amsterdam, Amsterdam, The Netherlands.
Cornelis P L
CP
Paul
Department of Neurosurgery, VU University Medical Center, Amsterdam, The Netherlands.
David P
DP
Noske
Department of Neurosurgery, VU University Medical Center, Amsterdam, The Netherlands.
W Peter
WP
Vandertop
Department of Neurosurgery, VU University Medical Center, Amsterdam, The Netherlands.|Neuroscience Center, Department of Neurology, Massachusetts General Hospital and Neuroscience Program, Harvard Medical School, Boston, MA, USA.
Thomas
T
Wurdinger
Department of Orthopedic Surgery, VU University Medical Center, MOVE Research Institute Amsterdam, Amsterdam, The Netherlands.
Marco N
MN
Helder
D000273:Adipose Tissue / Q000166:cytology*; D000818:Animals; D045744:Cell Line, Tumor; D002470:Cell Survival; D058948:Cell Tracking / Q000379:methods*; D005260:Female; D015870:Gene Expression; D017930:Genes, Reporter; D006041:Goats; D006801:Humans; D008163:Luminescent Measurements / Q000379:methods; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism; D009924:Organ Culture Techniques; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism*; D014161:Transduction, Genetic