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Crown Bioscience Inc.

5 ARTICLES PUBLISHED IN JoVE

Biology

Dopamine Release at Individual Presynaptic Terminals Visualized with FFNs

Hui Zhang^1,2, Niko G. Gubernator^3,4, Minerva Yue¹, Roland G. W. Staal¹, Eugene V. Mosharov¹, Daniela Pereira¹, Vojtech Balsanek³, Paul A. Vadola³, Bipasha Mukherjee⁵, Robert H. Edwards⁵, David Sulzer^1,2,6, Dalibor Sames³

¹Departments of Neurology, Columbia University, ²Departments of Psychiatry and Pharmacology, Columbia University, ³Department of Chemistry, Columbia University, ⁴eMolecules, Inc., ⁵Departments of Neurology and Physiology, University of California School of Medicine, San Francisco, ⁶Division of Molecular Therapeutics, New York Psychiatric Institute

A new means to measure neurotransmission optically using fluorescent dopamine analogs.

Cancer Research

Immunophenotyping of Orthotopic Homograft (Syngeneic) of Murine Primary KPC Pancreatic Ductal Adenocarcinoma by Flow Cytometry

Xiaoyu An^*1,2, Xuesong Ouyang^*1, Hui Zhang¹, Tingting Li¹, Yu-yang Huang¹, Zhiyuan Li¹, Demin Zhou², Qi-Xang Li^1,2

¹Crown Bioscience Inc., ²State Key Laboratory of Natural and Biomimetic Drugs, Peking University

The experimental procedure on the immunophenotyping of murine orthotopic PDAC homografts aims at profiling the tumor immuno-microenvironment. Tumors are orthotopically implanted via surgery. Tumors of 200–600 mm³ in size were harvested and dissociated to prepare single-cell suspensions, followed by multi-immune marker FACS analysis using different fluorescently-labeled antibodies.

Cancer Research

Creating Matched In vivo/In vitro Patient-Derived Model Pairs of PDX and PDX-Derived Organoids for Cancer Pharmacology Research

Xiaoxi Xu¹, Limei Shang¹, Philip Wang², Jun Zhou², Xuesong Ouyang², Meiling Zheng¹, Binchen Mao², Likun Zhang², Bonnie Chen¹, Jingjing Wang², Jing Chen³, Wubin Qian², Sheng Guo², Yujun Huang², Qi-Xiang Li³

¹Crown Bioscience Inc., Changping District, Beijing, China, ²Crown Bioscience Inc., Taicang, Jiangsu, China, ³Crown Bioscience Inc., San Diego, CA, USA

A method is described to create organoids using patient-derived xenografts (PDX) for in vitro screening, resulting in matched pairs of in vivo/in vitro models. PDX tumors were harvested/processed into small pieces mechanically or enzymatically, followed by the Clevers’ method to grow tumor organoids that were passaged, cryopreserved and characterized against the original PDX.

Cancer Research

Medium-Throughput Drug- and Radiotherapy Screening Assay using Patient-Derived Organoids

Marrit Putker^*1, Rosemary Millen^*2, René Overmeer^*3, Else Driehuis^2,4, Maurice M. J. M. Zandvliet⁵, Hans Clevers², Sylvia F. Boj³, Qi-Xiang Li⁶

¹Crown Bioscience Netherlands B.V., ²Oncode Institute, Hubrecht Institute, Royal Netherlands Academy of Arts and Sciences (KNAW) and University Medical Center Utrecht, ³Hubrecht Organoid Technology (HUB), ⁴Pathology dept, University Medical Center Utrecht, ⁵Department of Clinical Sciences - Companion Animals, Faculty of Veterinary Medicine, Utrecht University, ⁶Crown Bioscience Inc.

We describe detailed protocols to use patient-derived organoids for medium-throughput therapy sensitivity screenings. Therapies tested include chemotherapy, radiotherapy, and chemo-radiotherapy. Adenosine triphosphate levels are used as a functional readout.

Neuroscience

Measurement of Oxygen Consumption Rate in Acute Striatal Slices from Adult Mice

Lianteng Zhi¹, Jingyu Zhao¹, David Jaffe¹, Yuanxin Chen¹, Ninghan Wang^1,2, Qi Qin¹, Erin L. Seifert³, Chenjian Li⁴, Hui Zhang¹

¹Department of Neuroscience, Thomas Jefferson University, ²School of Biomedical Engineering, Drexel University, ³Department of Pathology, MitoCare Center, Thomas Jefferson University, ⁴School of Life Sciences, Peking University

Oxygen consumption rate (OCR) is a common proxy for mitochondrial function and can be used to study different disease models. We developed a new method using a Seahorse XF analyzer to directly measure the OCR in acute striatal slices from adult mice that is more physiologically relevant than other methods.