Chinese University of Hong Kong

9 ARTICLES PUBLISHED IN JoVE

Medicine

Evaluation of Cancer Stem Cell Migration Using Compartmentalizing Microfluidic Devices and Live Cell Imaging

Yu Huang^*1,2, Basheal Agrawal^*3, Paul A. Clark³, Justin C. Williams^1,2,3, John S. Kuo^3,4

¹Department of Biomedical Engineering, University of Wisconsin-Madison, ²Materials Science Program, University of Wisconsin-Madison, ³Department of Neurological Surgery, University of Wisconsin-Madison, ⁴Carbone Comprehensive Cancer Center and Center for Stem Cell and Regenerative Medicine, University of Wisconsin-Madison

A compartmentalizing microfluidic device for investigating cancer stem cell migration is described. This novel platform creates a viable cellular microenvironment and enables microscopic visualization of live cell locomotion. Highly motile cancer cells are isolated to study molecular mechanisms of aggressive infiltration, potentially leading to more effective future therapies.

Biology

Strategies for Tracking Anastasis, A Cell Survival Phenomenon that Reverses Apoptosis

Ho Lam Tang^1,2,3, Ho Man Tang^1,2,3, J. Marie Hardwick¹, Ming Chiu Fung²

¹W. Harry Feinstone Department of Molecular Microbiology and Immunology, Johns Hopkins University Bloomberg School of Public Health, ²School of Life Sciences, Chinese University of Hong Kong, ³Center for Cell Dynamics, Department of Biological Chemistry, Johns Hopkins University School of Medicine

The term anastasis refers to the phenomenon in which dying cells reverse a cell suicide process at a late stage, repair themselves, and ultimately survive. Here we demonstrate protocols for detecting and tracking cells that undergo anastasis.

Biology

In Vivo Biosensor Tracks Non-apoptotic Caspase Activity in Drosophila

Ho Lam Tang¹, Ho Man Tang¹, Ming Chiu Fung², J. Marie Hardwick¹

¹W. Harry Feinstone Department of Molecular Microbiology and Immunology, Johns Hopkins University Bloomberg School of Public Health, ²School of Life Sciences, Chinese University of Hong Kong

To detect healthy cells in whole animals that contain low levels of caspase activity, the highly sensitive biosensor designated CaspaseTracker was generated for Drosophila. Caspase-dependent biosensor activity is detected in long-lived healthy cells throughout the internal organs of adult animals reared under optimized conditions in the absence of death stimuli.

Medicine

Detecting Anastasis In Vivo by CaspaseTracker Biosensor

Ho Man Tang^1,2, Ming Chiu Fung², Ho Lam Tang³

¹Institute for Basic Biomedical Sciences, Johns Hopkins University School of Medicine, ²School of Life Sciences, Chinese University of Hong Kong, ³Department of Neurosurgery, Johns Hopkins University School of Medicine

Anastasis is technically challenging to detect in vivo because the cells that have reversed the cell death process can be morphologically indistinguishable from normal healthy cells. Here we describe protocols for detecting and tracking cells that undergo anastasis in live animals by using our newly developed in vivo CaspaseTracker biosensor system.

Developmental Biology

A 5-mC Dot Blot Assay Quantifying the DNA Methylation Level of Chondrocyte Dedifferentiation In Vitro

Zhaofeng Jia^1,2, Yujie Liang³, Bin Ma^4,5, Xiao Xu^2,6, Jianyi Xiong², Li Duan², Daping Wang^1,2

¹Guangzhou Medical University, ²Shenzhen Key Laboratory of Tissue Engineering, Shenzhen Laboratory of Digital Orthopeadic Engineering, Department of Orthopedics, Shenzhen Second People's Hospital (The First Hospital Affiliated to Shenzhen University), ³Department of Chemistry, The Chinese University of Hong Kong, ⁴School of Biomedical Engineering, Shanghai Jiao Tong University, ⁵Renji Hospital Clinical Stem Cell Research Center, Shanghai Jiao Tong University School of Medicine, ⁶Shantou University Medical College

We present a method to quantify DNA methylation based on the 5-methylcytosine (5-mC) dot blot. We determined the 5-mC levels during chondrocyte dedifferentiation. This simple technique could be used to quickly determine the chondrocyte phenotype in ACI treatment.

Biology

Magnetic-Activated Cell Sorting Strategies to Isolate and Purify Synovial Fluid-Derived Mesenchymal Stem Cells from a Rabbit Model

Zhaofeng Jia^1,2,3, Yujie Liang^4,5, Xingfu Li^2,3, Xiao Xu^1,2,3, Jianyi Xiong^2,3, Daping Wang^1,2,3, Li Duan^2,3

¹Postgraduate institution, Guangzhou Medical University, ²Guangdong Provincial Research Center for Artificial Intelligence and Digital Orthopedic Technology, ³Shenzhen Key Laboratory of Tissue Engineering, Shenzhen Laboratory of Digital Orthopaedic Engineering, Shenzhen Second People's Hospital (The First Hospital Affiliated to Shenzhen University), ⁴Department of Chemistry, Chinese University of Hong Kong, ⁵Shenzhen Kangning Hospital, Shenzhen Mental Health Center

This article presents a simple and economic protocol for the straightforward isolation and purification of mesenchymal stem cells from New Zealand white rabbit synovial fluid.

Developmental Biology

A Familial Hypercholesterolemia Human Liver Chimeric Mouse Model Using Induced Pluripotent Stem Cell-derived Hepatocytes

Jiayin Yang^*1,2,6, Lai-Yung Wong^*2, Xiao-Yu Tian^*3, Rui Wei^1,2, Wing-Hon Lai², Ka-Wing Au², Zhiwei Luo^4,5, Carl Ward^4,5, Wai-In Ho², David P. Ibañez^4,5, Hao Liu^4,5, Xichen Bao^4,5, Baoming Qin^4,5, Yu Huang³, Miguel A. Esteban^4,5,7, Hung-Fat Tse^1,2,6,7

¹Department of Medicine, University of Hong Kong-Shenzhen Hospital, ²The Cardiology Division, Department of Medicine, Li Ka Shing Faculty of Medicine, University of Hong Kong, ³School of Biomedical Sciences, Institute of Vascular Medicine, Li Ka Shing Institute of Health Sciences, Chinese University of Hong Kong, ⁴Key Laboratory of Regenerative Biology of the Chinese Academy of Sciences, Joint School of Life Sciences, Guangzhou Institutes of Biomedicine and Health and Guangzhou Medical University, ⁵Laboratory of RNA, Chromatin, and Human Disease, CAS Key Laboratory of Regenerative Biology and Guangdong Provincial Key Laboratory of Stem Cells and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, ⁶Research Centre of Heart, Brain, Hormone, and Healthy Ageing, Li Ka Shing Faculty of Medicine, University of Hong Kong, ⁷Hong Kong-Guangdong Stem Cell and Regenerative Medicine Research Centre, University of Hong Kong and Guangzhou Institutes of Biomedicine and Health

Here, we present a protocol to generate a human liver chimeric mouse model of familial hypercholesterolemia using human induced pluripotent stem cell-derived hepatocytes. This is a valuable model for testing new therapies for hypercholesterolemia.

Biology

Large-Scale Preparation of Synovial Fluid Mesenchymal Stem Cell-Derived Exosomes by 3D Bioreactor Culture

Li Duan^1,2, Xingfu Li^1,2, Xiao Xu^1,2, Limei Xu^1,2, Daping Wang^1,2, Kan Ouyang^1,2, Yujie Liang^1,3

¹Department of Orthopedics, The First affiliated hospital of Shenzhen University, Shenzhen Second People's Hospital, ²Guangdong Provincial Research Center for Artificial Intelligence and Digital Orthopedic Technology, Shenzhen Second People's Hospital, ³Department of Child and Adolescent Psychiatry, Shenzhen Kangning Hospital, Shenzhen Mental Health Center, Shenzhen Key Laboratory for Psychological Healthcare & Shenzhen Institute of Mental Health

Here, we present a protocol to produce a large number of GMP-grade exosomes from synovial fluid mesenchymal stem cells using a 3D bioreactor.

Biology

Visualization and Quantification of Brown and Beige Adipose Tissues in Mice using [¹⁸F]FDG Micro-PET/MR Imaging

Qing Liu^*1,2, Kel Vin Tan^*3, Hing-Chiu Chang³, Pek-Lan Khong³, Xiaoyan Hui^1,2,4

¹State Key Laboratory of Pharmaceutical Biotechnology, The University of Hong Kong, ²Department of Medicine, Li Ka Shing Faculty of Medicine, The University of Hong Kong, ³Department of Diagnostic Radiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, ⁴School of Biomedical Sciences, Institute of Vascular Medicine, Chinese University of Hong Kong

Functional imaging and quantitation of thermogenic adipose depots in mice using a micro-PET/MR imaging-based approach.