Homo sapiens
Protocols: Peripheral blood mononuclear cells (PBMCs) were isolated from whole blood with SepMate isolation tubes and Lymphoprep density gradient. PBMCs were then incubated with the following panel of antibodies; APC anti-human CD14 (Biolegend: 301807), Brilliant Violet 421 anti-human CD16 (Biolegend: 302037), PE anti-human CD66 (Biolegend: 342303), and PE/Cy7 anti-human HLA (Biolegend: 327017). Dead cells were excluded by staining with propidium iodide (Biolegend:421301). Flow cytometry and cell sorting was performed using a Beckman Coulter Astrios EQ cell sorter. Data were analysed with Kaluza Analysis software. Total RNA was extracted from cell suspensions of classical monocytes (CD14++/CD16-/CD66b-/HLA-DR+) and intermediate monocytes (CD14++/CD16+/CD66b-/HLA-DR+) using TRIzol reagent. Next generation RNA sequencing was performed by Genewiz. SMART-Seq v4 Ultra Low Input Kit for Sequencing was used for full-length cDNA synthesis and amplification (Clontech, Mountain View, CA), and Illumina Nextera XT library was used for sequencing library preparation. Briefly, cDNA was fragmented, and adaptor was added using Transposase, followed by limited-cycle PCR to enrich and add index to the cDNA fragments. The final library was assessed with Agilent TapeStation.
disease
osteoarthritis, knee
individual
WX123
organism
Homo sapiens
organism
Homo sapiens
body mass index
28.6
collection date
not collected
sex
Female
scientific_name
Homo sapiens
common name
human
organism part
Intermediate Monocyte
developmental stage
adult
kellgren-lawrence radiographic knee oa score
1
isolate
not applicable
numerical rating pain scale score
4
age
60
geographic location (country and/or sea)
not collected