University of Massachusetts Chan Medical School

3 ARTICLES PUBLISHED IN JoVE

Neuroscience

Human Microglia-like Cells: Differentiation from Induced Pluripotent Stem Cells and In Vitro Live-cell Phagocytosis Assay using Human Synaptosomes

Salome Funes^1,2, Daryl A. Bosco¹

¹Department of Neurology, University of Massachusetts Chan Medical School, ²Translation Science Program, Morningside Graduate School of Biomedical Sciences, University of Massachusetts Chan Medical School

This protocol describes the differentiation process of human induced pluripotent stem cells (iPSCs) into microglia-like cells for in vitro experimentation. We also include a detailed procedure for generating human synaptosomes from iPSC-derived lower motor neurons that can be used as a substrate for in vitro phagocytosis assays using live-cell imaging systems.

Neuroscience

Intravital Imaging of Fluorescent Protein Expression in Mice with a Closed-Skull Traumatic Brain Injury and Cranial Window Using a Two-Photon Microscope

Jianjun Zhong^1,2, Georgia Gunner³, Nils Henninger¹, Dorothy P. Schafer³, Daryl A. Bosco¹

¹Department of Neurology, University of Massachusetts Chan Medical School, ²Department of Neurosurgery, The First Affiliated Hospital of Chongqing Medical University, ³Department of Neurobiology, University of Massachusetts Chan Medical School

This study demonstrates delivery of a repetitive traumatic brain injury to mice and simultaneous implantation of a cranial window for subsequent intravital imaging of a neuron-expressed EGFP using two-photon microscopy.

Biochemistry

Oxygen-Independent Assays to Measure Mitochondrial Function in Mammals

Tenzin Tseyang^*1, Jonathan Valeros^*1, Paula Vo^*1, Jessica B. Spinelli^1,2

¹Program in Molecular Medicine, University of Massachusetts Chan Medical School, ²Cancer Center, University of Massachusetts Chan Medical School

Here, we present a compilation of assays to directly measure mitochondrial function in mammalian cells independently of their ability to consume molecular oxygen.