Title | Specimen Preparation for Single-Cell Sequencing Analysis of Skeletal Cells. |
Publication Type | Journal Article |
Year of Publication | 2021 |
Authors | Debnath S, Greenblatt MB |
Journal | Methods Mol Biol |
Volume | 2221 |
Pagination | 89-100 |
Date Published | 2021 |
ISSN | 1940-6029 |
Keywords | Animals, Bone and Bones, Mice, Osteocytes, Single-Cell Analysis, Specimen Handling |
Abstract | Recent work emphasizes that bone comprises numerous mesenchymal cell types each with different biologic functions, and deconvoluting the functions of these cells requires technical approaches with single-cell resolution, such as single-cell RNA sequencing (scRNA-seq). A critical step in conducting a successful single-cell sequencing study of bone is generation of a single-cell suspension of skeletal cells while preserving cell viability. Here we describe a method to prepare single-cell suspensions from skeletal tissue in preparation for single-cell sequencing studies. Also included are optional steps for fluorescence-activated cell sorting (FACS) of skeletal cells, which allows subsequent scRNA-seq studies to be focused on specific populations of interest. |
DOI | 10.1007/978-1-0716-0989-7_7 |
Alternate Journal | Methods Mol Biol |
PubMed ID | 32979200 |
Related Faculty:
Matthew B. Greenblatt, M.D., Ph.D.