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High throughput imaging identifies a spatially localized response of primary fetal pulmonary artery endothelial cells to insulin-like growth factor 1 treatment

Christina Kim, Gregory J Seedorf, Steven H Abman, View ORCID ProfileDouglas P Shepherd
doi: https://doi.org/10.1101/674499
Christina Kim
aDepartment of Surgery, University of Colorado Anschutz Medical Campus, Aurora, CO 80045
bPediatric Heart Lung Center, University of Colorado Anschutz Medical Campus, Aurora, CO 80045
dDepartment of Pharmacology, University of Colorado Anschutz Medical Campus, Aurora, CO 80045
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Gregory J Seedorf
bPediatric Heart Lung Center, University of Colorado Anschutz Medical Campus, Aurora, CO 80045
cDepartment of Pediatrics, University of Colorado Anschutz Medical Campus, Aurora, CO 80045
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Steven H Abman
bPediatric Heart Lung Center, University of Colorado Anschutz Medical Campus, Aurora, CO 80045
cDepartment of Pediatrics, University of Colorado Anschutz Medical Campus, Aurora, CO 80045
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Douglas P Shepherd
bPediatric Heart Lung Center, University of Colorado Anschutz Medical Campus, Aurora, CO 80045
dDepartment of Pharmacology, University of Colorado Anschutz Medical Campus, Aurora, CO 80045
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  • ORCID record for Douglas P Shepherd
  • For correspondence: douglas.shepherd@asu.edu
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Abstract

A common strategy to measure the efficacy of drug treatment is the in vitro comparison of ensemble readouts with and without treatment, such as proliferation and cell death. A fundamental assumption underlying this approach is there is minimal cell to cell variability in the response to drug. Here, we demonstrate that ensemble and non-spatial single cell readouts applied to primary cells lead to incomplete conclusions due to cell to cell variability. We exposed primary fetal pulmonary artery endothelial cells (PAEC) isolated from healthy newborn healthy and persistent pulmonary hypertension of the newborn (PPHN) sheep to the growth hormone insulin-like growth factor 1 (IGF-1). We found that IGF-1 increased proliferation and branch points in tube formation assays but not angiogenic signaling proteins at the population level for both cell types. We hypothesized that this molecular ambiguity was due to the presence of cellular subpopulations with variable responses to IGF-1. Using high throughput single cell imaging, we discovered a spatially localized response to IGF-1. This suggests localized signaling or heritable cell response to external stimuli may ultimately be responsible for our observations. Discovering and further exploring these rare cells is critical to finding new molecular targets to restore cellular function.

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  • https://zenodo.org/record/2803235#.XQgJxlxKg2w

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted June 17, 2019.
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High throughput imaging identifies a spatially localized response of primary fetal pulmonary artery endothelial cells to insulin-like growth factor 1 treatment
Christina Kim, Gregory J Seedorf, Steven H Abman, Douglas P Shepherd
bioRxiv 674499; doi: https://doi.org/10.1101/674499
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High throughput imaging identifies a spatially localized response of primary fetal pulmonary artery endothelial cells to insulin-like growth factor 1 treatment
Christina Kim, Gregory J Seedorf, Steven H Abman, Douglas P Shepherd
bioRxiv 674499; doi: https://doi.org/10.1101/674499

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