A 3D biofabricated cutaneous squamous cell carcinoma tissue model with multi-channel confocal microscopy imaging biomarkers to quantify antitumor effects of chemotherapeutics in tissue
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James R. Browning1, Paige Derr2, Kristy Derr2, Nicole Doudican3, Sam Michael2, Samantha R. Lish1, Nicholas A. Taylor3, James G. Krueger1, Marc Ferrer2, John A. Carucci3 and Daniel S. Gareau1
1 Laboratory for Investigative Dermatology, The Rockefeller University, New York, New York, USA
2 National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, Maryland, USA
3 The Ronald O. Perelman Department of Dermatology, New York University School of Medicine, New York, New York, USA
|Daniel S. Gareau,||email:||[email protected]|
Keywords: squamous cell carcinoma; screening; 3D printing; in vitro model; confocal microscopy
Received: January 05, 2020 Accepted: April 03, 2020 Published: July 07, 2020
Cutaneous squamous cell carcinoma (cSCC) causes approximately 10,000 deaths annually in the U. S. Current therapies are largely ineffective against metastatic and locally advanced cSCC. There is a need to identify novel, effective, and less toxic small molecule cSCC therapeutics. We developed a 3-dimensional bioprinted skin (3DBPS) model of cSCC tumors together with a microscopy assay to test chemotherapeutic effects in tissue. The full thickness SCC tissue model was validated using hematoxylin and eosin (H&E) and immunohistochemical histological staining, confocal microscopy, and cDNA microarray analysis. A nondestructive, 3D fluorescence confocal imaging assay with tdTomato-labeled A431 SCC and ZsGreen-labeled keratinocytes was developed to test efficacy and general toxicity of chemotherapeutics. Fluorescence-derived imaging biomarkers indicated that 50% of cancer cells were killed in the tissue after 1?M 5-Fluorouracil 48-hour treatment, compared to a baseline of 12% for untreated controls. The imaging biomarkers also showed that normal keratinocytes were less affected by treatment (11% killed) than the untreated tissue, which had no significant killing effect. Data showed that 5-Fluorouracil selectively killed cSCC cells more than keratinocytes. Our 3DBPS assay platform provides cellular-level measurement of cell viability and can be adapted to achieve nondestructive high-throughput screening (HTS) in bio-fabricated tissues.
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