Open AccessReal-Time Imaging of CCL5-Induced Migration of Periosteal Skeletal Stem Cells in Mice
Author(s) -
Laura Ortinau,
Kevin Lei,
Youngjae Jeong,
Dongsu Park
Publication year - 2020
Publication title -
journal of visualized experiments
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.596
H-Index - 91
ISSN - 1940-087X
DOI - 10.3791/61162
Subject(s) - calvaria , mesenchymal stem cell , stem cell , microbiology and biotechnology , bone healing , ccl5 , intravital microscopy , cell migration , periosteum , medicine , pathology , cell , chemistry , biology , anatomy , immunology , t cell , in vitro , immune system , biochemistry , il 2 receptor , microcirculation
Periosteal skeletal stem cells (P-SSCs) are essential for lifelong bone maintenance and repair, making them an ideal focus for the development of therapies to enhance fracture healing. Periosteal cells rapidly migrate to an injury to supply new chondrocytes and osteoblasts for fracture healing. Traditionally, the efficacy of a cytokine to induce cell migration has only been conducted in vitro by performing a transwell or scratch assay. With advancements in intravital microscopy using multiphoton excitation, it was recently discovered that 1) P-SSCs express the migratory gene CCR5 and 2) treatment with the CCR5 ligand known as CCL5 improves fracture healing and the migration of P-SSCs in response to CCL5. These results have been captured in real-time. Described here is a protocol to visualize P-SSC migration from the calvarial suture skeletal stem cell (SSC) niche towards an injury after treatment with CCL5. The protocol details the construction of a mouse restraint and imaging mount, surgical preparation of the mouse calvaria, induction of a calvaria defect, and acquisition of time-lapse imaging.