CHU de Québec Research Center

4 ARTICLES PUBLISHED IN JoVE

Developmental Biology

Generation of Human Adipose Stem Cells through Dedifferentiation of Mature Adipocytes in Ceiling Cultures

Julie Lessard¹, Julie Anne Côté¹, Marc Lapointe¹, Mélissa Pelletier², Mélanie Nadeau¹, Simon Marceau¹, Laurent Biertho¹, André Tchernof^1,2,3

¹IUCPQ Research Center, ²CHU de Québec Research Center, ³Laval University

Mature adipocytes may represent an abundant source of stem cells through dedifferentiation, which leads to a homogenous population of fibroblast-like cells. Collagenase digestion is used to isolate mature adipocytes from human fat. The goal of our protocol is to obtain multipotent, dedifferentiated fat cells from human mature adipocytes.

Neuroscience

Correlative Light and Electron Microscopy to Study Microglial Interactions with β-Amyloid Plaques

Kanchan Bisht^*1, Hassan El Hajj^*1, Julie C. Savage¹, Maria G. Sánchez¹, Marie-Ève Tremblay¹

¹Neurosciences Axis, CHU de Québec Research Center

This article describes a protocol for visualizing amyloid Aβ plaques in Alzheimer's disease mouse models using methoxy-X04, which crosses the blood-brain barrier and selectively binds to β-pleated sheets found in dense core Aβ plaques. It allows pre-screening of plaque-containing tissue sections prior to immunostaining and processing for electron microscopy.

Immunology and Infection

Immunofluorescence Staining Using IBA1 and TMEM119 for Microglial Density, Morphology and Peripheral Myeloid Cell Infiltration Analysis in Mouse Brain

Fernando González Ibanez^1,2, Katherine Picard^1,2, Maude Bordeleau^1,3, Kaushik Sharma^1,2,4, Kanchan Bisht^1,2,4, Marie-Ève Tremblay^1,2

¹Axe Neurosciences, Centre de Recherche du CHU de Québec-Université Laval, ²Département de Médecine Moléculaire, Faculté de Médecine, Université Laval, ³Integrated Program in Neuroscience, McGill University, ⁴Center for Brain Immunology and Glia (BIG), University of Virginia

This protocol describes a step-by-step workflow for immunofluorescent costaining of IBA1 and TMEM119, in addition to analysis of microglial density, distribution, and morphology, as well as peripheral myeloid cell infiltration in mouse brain tissue.

Immunology and Infection

Precise Brain Mapping to Perform Repetitive In Vivo Imaging of Neuro-Immune Dynamics in Mice

Kanchan Bisht^1,2, Kaushik Sharma^1,2, Ukpong B. Eyo^1,2

¹Center for Brain Immunology and Glia (BIG), University of Virginia, ²Department of Neuroscience, University of Virginia

This protocol describes a chronic cranial window implantation technique that can be used for longitudinal imaging of neuro-glio-vascular structures, interactions, and function in both healthy and diseased conditions. It serves as a complementary alternative to the transcranial imaging approach that, while often preferred, possesses some critical limitations.