A Comparison of The Efficiency of RNA Extractionfrom Extracellular Vesicles of Endometriosis Patients UsingTwo Different Methods

Background: Extracellular vehicles (EVs) involved in multiplephysiological processes and are major contributors throughprogression of disease. As EVs contain lipoproteins, DNA,mRNA and are present in body fluids, are good candidate fors tudying diseases by minimally invasive methods. Their role ina variety of diseases; has been widely s tudied but little is knownabout their role in endometriosis. Mos t gene expression s tudiesof endometriosis are performed on endometrial tissue, whichhas an invasive sampling process. Our main goal is to s tudy ofgene expression in endometriosis using EVs which are presentin the circulation ins tead of endometriotic tissues.Materials and Methods: So, we optimized EV isolation fromplasma of endometriosis patients and compared two methods ofRNA extraction from EVs. We conducted a pilot s tudy based onethical approval on blood samples of ۴ endometriosis womenreferred to Royan Ins titute after consent was obtained fromthem. Plasma was divided into two parts. Firs t part was used toseparate EVs using ultracentrifuge followed by characterizationby morphology, DSL and wes tern blot (CD۸۱, CD۹, TSG۱۰۱).Characterized EVs and second part of plasma (without furthermanipulation) were used to RNA extraction using Plasma/serum circulating and exosomal RNA purification kit. AftercDNA synthesis the expression of EN gene (which is highlyexpressed in endometriosis) evaluated quantitatively by realtimePCR.Results: The results showed that direct extraction of EV RNAsfrom plasma is more efficient than RNA extraction from isolatedEVs using ultracentrifuge, and yielded lower CTs throughreal-time PCR.Conclusion: These data imply the importance of selectin of theappropriate technique in the s tudy of EVs as small and delicatesamples that eliminate the need for invasive methods