|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|
|Keywords||Animals, Bone and Bones, Mice, Osteocytes, Single-Cell Analysis, Specimen Handling|
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.
|Alternate Journal||Methods Mol Biol|
Matthew B. Greenblatt, M.D., Ph.D.