Profiling the MicroRNA Payload of Exosomes Derived from Ex Vivo Primary Colorectal Fibroblasts.
The tumor microenvironment is a heterogeneous and dynamic network that exists between cancer and stroma, playing a critical role in cancer progression. Certain tumorigenic signals such as microRNAs are derived from the stroma and conveyed to cancer cells (and vice versa) in nanoparticles called exosomes. Their identification and characterization is an important step in better understanding cellular cross talk and its consequences. To this end we describe how to culture primary ex vivo derived fibroblasts from colorectal tissue, isolate their exosomes, extract exosomal RNA and perform microRNA profiling.
Cancer Sciences, University of Southampton, Somers Building, Tremona Road, Southampton, SO16 6YD, UK. r.bhome@soton.ac.uk.|University Surgical Unit, Southampton General Hospital, Tremona Road, Southampton, SO16 6YD, UK. r.bhome@soton.ac.uk.
Rahul
R
Bhome
Cancer Sciences, University of Southampton, Somers Building, Tremona Road, Southampton, SO16 6YD, UK.
Rebecca
R
Goh
Cancer Sciences, University of Southampton, Somers Building, Tremona Road, Southampton, SO16 6YD, UK.
Karen
K
Pickard
Cancer Sciences, University of Southampton, Somers Building, Tremona Road, Southampton, SO16 6YD, UK.
Massimiliano
M
Mellone
Cancer Sciences, University of Southampton, Somers Building, Tremona Road, Southampton, SO16 6YD, UK.
A Emre
AE
Sayan
Cancer Sciences, University of Southampton, Somers Building, Tremona Road, Southampton, SO16 6YD, UK.|University Surgical Unit, Southampton General Hospital, Tremona Road, Southampton, SO16 6YD, UK.
Alex
A
Mirnezami
D014408:Biomarkers, Tumor / Q000235:genetics / Q000378:metabolism; D002478:Cells, Cultured; D003106:Colon / Q000378:metabolism / Q000473:pathology; D015179:Colorectal Neoplasms / Q000235:genetics / Q000378:metabolism*; D055354:Exosomes / Q000235:genetics / Q000378:metabolism*; D005347:Fibroblasts / Q000378:metabolism*; D020869:Gene Expression Profiling; D006801:Humans; D035683:MicroRNAs / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D061251:Primary Cell Culture; D060888:Real-Time Polymerase Chain Reaction; D012007:Rectum / Q000378:metabolism / Q000473:pathology
D014408:Biomarkers, Tumor; D035683:MicroRNAs
Colorectal cancer; Exosome; Fibroblast; Stroma; Tumor microenvironment