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University of Rochester, School of Medicine and Dentistry

5 ARTICLES PUBLISHED IN JoVE

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Genetics

Differentiation, Maintenance, and Analysis of Human Retinal Pigment Epithelium Cells: A Disease-in-a-dish Model for BEST1 Mutations
Alec Kittredge 1, Changyi Ji 1, Yu Zhang  1, Tingting Yang 1
1Department of Pharmacology and Physiology, University of Rochester, School of Medicine and Dentistry

Here we present a protocol to differentiate retinal pigment epithelium (RPE) cells from human pluripotent stem cells bearing patient-derived mutations. The mutant cell lines may be used for functional analyses including immunoblotting, immunofluorescence, and patch clamp. This disease-in-a-dish approach circumvents the difficulty of obtaining native human RPE cells.

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Biochemistry

Expression and Purification of Mammalian Bestrophin Ion Channels
Alec Kittredge 1, Nancy Ward 1, Austin Hopiavuori 1, Yu Zhang  1, Tingting Yang 1
1Department of Pharmacology and Physiology, University of Rochester, School of Medicine and Dentistry

The purification of ion channels is often challenging, but once achieved, it can potentially allow in vitro investigations of the functions and structures of the channels. Here, we describe the stepwise procedures for the expression and purification of mammalian bestrophin proteins, a family of Ca2+-activated Cl- channels.

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Medicine

Polyethyleneimine-coated Iron Oxide Nanoparticles as a Vehicle for the Delivery of Small Interfering RNA to Macrophages In Vitro and In Vivo
Na Jia *1, Haoan Wu *2, Juanli Duan 1, Chunxue Wei 1, Kaixuan Wang 2, Yu Zhang  2, Xiaohua Mao 1
1Key Laboratory of Ministry of Education for Developmental Genes & Human Diseases, Institute of Life Sciences and School of Medicine, Southeast University, 2State Key Laboratory of Bioelectronics, Jiangsu Key Laboratory for Biomaterials & Devices, School of Biological Science & Medical Engineering, Southeast University

We describe a method of using polyethyleneimine (PEI)-coated superparamagnetic iron oxide nanoparticles for transfecting macrophages with siRNA. These nanoparticles can efficiently deliver siRNA to macrophages in vitro and in vivo and silence target gene expression.

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JoVE Journal

Using Enhanced Green Fluorescence Protein-expressing Escherichia Coli to Assess Mouse Peritoneal Macrophage Phagocytosis
Yu Zhang  *1, Guan Wang *1, Jia Lu 1, Li-ming Xu 2, Jun-yu Xiong 1
1Anesthesiology Department, The Second Hospital of Dalian Medical University, 2Scientific Research Center, The Second Hospital of Dalian Medical University

Here, we present a protocol to assess mouse peritoneal macrophage phagocytosis using enhanced green fluorescence protein-expressing Escherichia coli.

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Chemistry

Reactive Inkjet Printing and Propulsion Analysis of Silk-based Self-propelled Micro-stirrers
David A. Gregory 1, Piyush Kumar 1, Ana Jimenez-Franco 1, Yi Zhang 1, Yu Zhang  2, Stephen J. Ebbens 1, Xiubo Zhao 1
1Department of Chemical and Biological Engineering, University of Sheffield, 2School of Engineering, University of Liverpool

This protocol demonstrates the ability to utilize reactive inkjet printing to print self-motile biocompatible and environmentally friendly micro-stirrers for use in biomedical and environmental applications.

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