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3D Bioprinted Intrahepatic Cholangiocarcinoma Model and 7-Day Perfusion with Gemcitabine and Cisplatin: Modeling the Tumor Microenvironment and Response to Chemotherapy

A. M. Chen1, W. Zhang1, J. Sterner2, L. Kennedy3, A. Isidan1, E. Gramelspacher1, H. L. Francis3, G. Alpini3, L. J. Smith2, P. Li1, B. Ekser1

1Division of Transplant Surgery - Indiana University, Indianapolis, IN, 23D Bioprinting Core and Department of Radiology and Imaging Sciences - Indiana University, Indianapolis, IN, 3Division of Gastroenterology, Richard L. Roudebush VA Medical Center and Indiana University, Indianapolis, IN

Meeting: 2020 American Transplant Congress

Abstract number: D-307

Keywords: Apoptosis, Bile duct, Bioengineering, Liver

Session Information

Date: Saturday, May 30, 2020

Session Name: Poster Session D: Cellular Therapies, Tissue Engineering / Regenerative Medicine

Session Time: 3:15pm-4:00pm

 Presentation Time: 3:30pm-4:00pm

Location: Virtual

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*Purpose: Over 8,000 patients are newly diagnosed with cholangiocarcinoma (CCA) in the United States each year.Understanding the CCA microenvironment will facilitate the development of novel therapeutic strategies and drug regimens. 3D-bioprinting is useful for studying cell-cell interaction and pathophysiological functions, with advantages over traditional 2D cell culture. We developed and tested a novelscaffold-free3D-bioprinted intrahepatic CCA model, which was continuously perfused with gemcitabine and cisplatin to elucidate tumor-support cell interactions in the setting of chemotherapy.

*Methods: All humanprimary intrahepatic CCA (HuCC-T1), hepatic stellate cells (HSC), and cholangiocytes (CHO) were co-cultured to form spheroids. Spheroids were bioprinted using the Regenova 3D-Bioprinter into liver tissue constructs, as HSC-only (control) or CCA:HSC:CHO (2:1:1) combination. Constructs were perfused continuously for 7 days with media-alone or media-supplemented-with-gemcitabine-and-cisplatin (10 μM/L and 20 μM/L, respectively) in the FABRICA bioreactor. Real-time PCR was used to analyze the transcription of AFP, BAX, COL1A1, TGFβ1 for functionality of cancer/progenitor cell markers, apoptosis, extracellular matrix (ECM) deposition, and metastatic potential in spheroids/constructs on days 0, 1, 7.

*Results: The 3D-CCA model was successfully bioprinted and perfused with media-alone or media-gemcitabine/cisplatin for 7 days. Gemcitabine and cisplatin prevented ECM formation and robust tissue fusion in chemotherapy-treated constructs (Figure 1B). Overall, AFP, BAX, COL1A1, TGFβ1 gene expression increased over the treatment period (Figure 1C). Gemcitabine/cisplatin treatment was associated at day 7 with markedly elevated AFP expression, decreasing COL1A1 over time, higher BAX in combined conditions, and higher TGFβ1 expression in HSC-containing constructs compared to spheroids.

*Conclusions: Development of an inexpensive and efficient 3D-CCA model enables us to study its microenvironment for expression of cancer/progenitor markers, apoptosis, collagen formation, and invasion/metastasis under chemotherapy.

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To cite this abstract in AMA style:

Chen AM, Zhang W, Sterner J, Kennedy L, Isidan A, Gramelspacher E, Francis HL, Alpini G, Smith LJ, Li P, Ekser B. 3D Bioprinted Intrahepatic Cholangiocarcinoma Model and 7-Day Perfusion with Gemcitabine and Cisplatin: Modeling the Tumor Microenvironment and Response to Chemotherapy [abstract]. Am J Transplant. 2020; 20 (suppl 3). https://atcmeetingabstracts.com/abstract/3d-bioprinted-intrahepatic-cholangiocarcinoma-model-and-7-day-perfusion-with-gemcitabine-and-cisplatin-modeling-the-tumor-microenvironment-and-response-to-chemotherapy/. Accessed January 16, 2021.

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