Johns Hopkins Medical Institute

4 ARTICLES PUBLISHED IN JoVE

Immunology and Infection

Murine Hind Limb Long Bone Dissection and Bone Marrow Isolation

Sarah R. Amend¹, Kenneth C. Valkenburg¹, Kenneth J. Pienta¹

¹Department of Urology, Johns Hopkins University

Here we present a protocol for the dissection of hind limb long bones (femurs and tibiae) from the laboratory mouse. We further describe a rapid technique for bone marrow isolation from these bones that utilizes centrifugation for removal of bone marrow from the bone marrow space.

Medicine

Murine Prostate Micro-dissection and Surgical Castration

Kenneth C. Valkenburg¹, Sarah R. Amend¹, Kenneth J. Pienta¹

¹Department of Urology, Johns Hopkins University

This manuscript describes the protocols for prostate micro-dissection and surgical castration in the laboratory mouse. We also depict representative results produced by these protocols. Finally, we discuss the advantages and utilization of these protocols.

Bioengineering

Generation of Heterogeneous Drug Gradients Across Cancer Populations on a Microfluidic Evolution Accelerator for Real-Time Observation

Ke-Chih Lin¹, Gonzalo Torga², Yusha Sun¹, Kenneth J. Pienta², James C. Sturm¹, Robert H. Austin¹

¹Princeton University, ²Johns Hopkins Medical Institute

We present a microfluidic cancer-on-chip model, the "Evolution Accelerator" technology, which provides a controllable platform for long-term real-time quantitative studies of cancer dynamics within well-defined environmental conditions at the single-cell level. This technology is expected to work as an in vitro model for fundamental research or pre-clinical drug development.

Biology

Extracellular Vesicle Uptake Assay via Confocal Microscope Imaging Analysis

Chi-Ju Kim^*1,2, Morgan D. Kuczler^*1, Liang Dong^1,3, Junyoung Kim^2,4, Sarah R. Amend¹, Yoon-Kyoung Cho^2,4, Kenneth J. Pienta¹

¹The Brady Urological Institute, Johns Hopkins University School of Medicine, ²Department of Biomedical Engineering, School of Life Sciences, Ulsan National Institute of Science and Technology (UNIST), ³Department of Urology, Renji Hospital, Shanghai Jiao Tong University School of Medicine, ⁴Center for Soft and Living Matter, Institute for Basic Science (IBS)

Extracellular vesicles (EVs) contribute to cellular biology and intercellular communications. There is a need for practical assays to visualize and quantify EVs uptake by the cells. The current protocol proposes the EV uptake assay by utilizing three-dimensional fluorescence imaging via confocal microscopy, following EV isolation by a nano-filtration-based microfluidic device.