Single cell RNA-sequencing analysis of human nonunion tissue
10X Genomics single cell RNA-sequencing of freshly isolated nonunion tissue from 63-year old male was conducted to interrogate non-hematopoietic tissue composition. The goal was to generate a dataset that can be compared with publicly available datasets of normal fracture tissue and cells associated with bone healing. Overall design: Nonunion tissue was harvested during corrective surgery 300 days after initial injury. After mechanical and enzymatic preparation, the single cell solution was stained for flow cytometry and 3x10 6 PI-CD45-CD235a- cells were sorted into a collection tube containing FACS buffer. Cells were then processed with 10X Chromium Next GEM Single Cell 3' GEM kit (10X Genomics Inc, v3.1) according to manufacturer's instruction to target 10,000 cells. A library ('Nonunion') was generated for partial lane sequencing on a NovaSeq6000.