title: ÐÐµÑÐ¾Ð´Ñ Ð²ÑÐ´ÐµÐ»ÐµÐ½Ð¸Ñ Ð¸ Ð°Ð½Ð°Ð»Ð¸Ð·Ð° ÑÐ¿ÐµÑÐ¸ÑÐ¸ÑÐµÑÐºÐ¸Ñ ÑÑÐ°ÐºÑÐ¸Ð¹ Ð½Ð°Ð½Ð¾Ð²ÐµÐ·Ð¸ÐºÑÐ» Ð¿Ð»Ð°Ð·Ð¼Ñ
title: Ð²ÑÐ¿ÑÑÐºÐ½Ð°Ñ ÐºÐ²Ð°Ð»Ð¸ÑÐ¸ÐºÐ°ÑÐ¸Ð¾Ð½Ð½Ð°Ñ ÑÐ°Ð±Ð¾ÑÐ° Ð¼Ð°Ð³Ð¸ÑÑÑÐ°
titleType: Subtitle
title: Methods for isolation and analysis of specific fractions of plasma nanovesicles
Creators: name: ÐÐ°Ð±ÐµÐ³Ð¸Ð½Ð°, ÐÐ¸Ð´Ð¸Ñ
Publisher: name: Ð¡Ð°Ð½ÐºÑ-ÐÐµÑÐµÑÐ±ÑÑÐ³ÑÐºÐ¸Ð¹ Ð¿Ð¾Ð»Ð¸ÑÐµÑÐ½Ð¸ÑÐµÑÐºÐ¸Ð¹ ÑÐ½Ð¸Ð²ÐµÑÑÐ¸ÑÐµÑ ÐÐµÑÑÐ° ÐÐµÐ»Ð¸ÐºÐ¾Ð³Ð¾
Year: 2021
Subjects: ÐÐ¸ÐºÑÐ¾Ð±Ð¸Ð¾Ð»Ð¾Ð³Ð¸Ñ
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description: ÐÐ°Ð½Ð½Ð°Ñ ÑÐ°Ð±Ð¾ÑÐ° Ð¿Ð¾ÑÐ²ÑÑÐµÐ½Ð° Ð¸ÑÑÐ»ÐµÐ´Ð¾Ð²Ð°Ð½Ð¸Ñ Ð¿ÐµÑÑÐ¿ÐµÐºÑÐ¸Ð² Ð¸ÑÐ¿Ð¾Ð»ÑÐ·Ð¾Ð²Ð°Ð½Ð¸Ñ Ð¼Ð¸ÐºÑÐ¾ Ð ÐÐ Ð¸Ð· ÑÐ¿ÐµÑÐ¸ÑÐ¸ÑÐµÑÐºÐ¸Ñ ÑÑÐ°ÐºÑÐ¸Ð¹ Ð²Ð½ÐµÐºÐ»ÐµÑÐ¾ÑÐ½ÑÑ Ð½Ð°Ð½Ð¾Ð²ÐµÐ·Ð¸ÐºÑÐ» (ÐÐÐ) Ð¿Ð»Ð°Ð·Ð¼Ñ Ð² ÐºÐ°ÑÐµÑÑÐ²Ðµ Ð¼Ð°ÑÐºÐµÑÐ¾Ð² Ð´Ð»Ñ Ð´Ð¸ÑÑÐµÑÐµÐ½ÑÐ¸Ð°Ð»ÑÐ½Ð¾Ð¹ Ð´Ð¸Ð°Ð³Ð½Ð¾ÑÑÐ¸ÐºÐ¸ Ð¾Ð½ÐºÐ¾Ð»Ð¾Ð³Ð¸ÑÐµÑÐºÐ¸Ñ Ð·Ð°Ð±Ð¾Ð»ÐµÐ²Ð°Ð½Ð¸Ð¹. ÐÐ°Ð´Ð°ÑÐ¸, ÐºÐ¾ÑÐ¾ÑÑÐµ ÑÐµÑÐ°Ð»Ð¸ÑÑ Ð² ÑÐ¾Ð´Ðµ Ð¸ÑÑÐ»ÐµÐ´Ð¾Ð²Ð°Ð½Ð¸Ñ: 1. ÐÑÐ´ÐµÐ»ÐµÐ½Ð¸Ðµ Ð¸ Ð°Ð½Ð°Ð»Ð¸Ð· Ð¾Ð±ÑÐµÐ¹ Ð¿Ð¾Ð¿ÑÐ»ÑÑÐ¸Ð¸ Ð½Ð°Ð½Ð¾Ð²ÐµÐ·Ð¸ÐºÑÐ». 2. Ð Ð°Ð·ÑÐ°Ð±Ð¾ÑÐºÐ° Ð¼ÐµÑÐ¾Ð´Ð° Ð´Ð»Ñ Ð²ÑÐ´ÐµÐ»ÐµÐ½Ð¸Ñ ÑÐ¿ÐµÑÐ¸ÑÐ¸ÑÐµÑÐºÐ¸Ñ ÑÑÐ°ÐºÑÐ¸Ð¹ Ð½Ð°Ð½Ð¾Ð²ÐµÐ·Ð¸ÐºÑÐ». 3. ÐÑÐ´ÐµÐ»ÐµÐ½Ð¸Ðµ Ð¢ÐÐ(+) Ð¸ ÐÐ¡ÐÐ(+) ÑÐ¿ÐµÑÐ¸ÑÐ¸ÑÐ½ÑÑ ÑÑÐ°ÐºÑÐ¸Ð¹ Ð½Ð°Ð½Ð¾Ð²ÐµÐ·Ð¸ÐºÑÐ». 4. ÐÑÐµÐ½ÐºÐ° Ð´Ð¸Ð°Ð³Ð½Ð¾ÑÑÐ¸ÑÐµÑÐºÐ¾Ð³Ð¾ Ð¿Ð¾ÑÐµÐ½ÑÐ¸Ð°Ð»Ð° Ð¸ÑÐ¿Ð¾Ð»ÑÐ·Ð¾Ð²Ð°Ð½Ð¸Ñ Ð¼Ð¸ÐºÑÐ¾Ð ÐÐ Ð¸Ð· Ð¢ÐÐ(+) Ð¸ ÐÐ¡ÐÐ(+) ÑÑÐ°ÐºÑÐ¸Ð¹ Ð½Ð°Ð½Ð¾Ð²ÐµÐ·Ð¸ÐºÑÐ». Ð Ð°Ð±Ð¾ÑÐ° Ð²ÑÐ¿Ð¾Ð»Ð½ÑÐ»Ð°ÑÑ Ð½Ð° Ð±Ð°Ð·Ðµ ÐÐÐÐ¦ Ð¾Ð½ÐºÐ¾Ð»Ð¾Ð³Ð¸Ð¸ Ð¸Ð¼ÐµÐ½Ð¸ Ð. Ð. ÐÐµÑÑÐ¾Ð²Ð°. ÐÐ±ÑÐ°Ñ Ð¿Ð¾Ð¿ÑÐ»ÑÑÐ¸Ñ Ð²ÐµÐ·Ð¸ÐºÑÐ» Ð±ÑÐ»Ð° Ð¿Ð¾Ð»ÑÑÐµÐ½Ð° Ð¼ÐµÑÐ¾Ð´Ð°Ð¼Ð¸ ÑÐ»ÑÑÑÐ°ÑÐµÐ½ÑÑÐ¸ÑÑÐ³Ð¸ÑÐ¾Ð²Ð°Ð½Ð¸Ñ, Ð´Ð²ÑÑÑÐ°Ð·Ð½Ð¾Ð¹ Ð¿Ð¾Ð»Ð¸Ð¼ÐµÑÐ½Ð¾Ð¹ ÑÐ¸ÑÑÐµÐ¼Ñ Ð¸ ÑÐºÑÐºÐ»ÑÐ·Ð¸Ð¾Ð½Ð½Ð¾Ð¹ ÑÑÐ¾Ð¼Ð°ÑÐ¾Ð³ÑÐ°ÑÐ¸Ð¸. Ð¡Ð¿ÐµÑÐ¸ÑÐ¸ÑÐµÑÐºÐ¸Ðµ ÑÑÐ°ÐºÑÐ¸Ð¸ Ð²ÐµÐ·Ð¸ÐºÑÐ» Ð²ÑÐ´ÐµÐ»ÑÐ»Ð¸ Ñ Ð¸ÑÐ¿Ð¾Ð»ÑÐ·Ð¾Ð²Ð°Ð½Ð¸ÐµÐ¼ Ð¼ÐµÑÐ¾Ð´Ð¸Ðº, Ð¾ÑÐ½Ð¾Ð²Ð°Ð½Ð½ÑÑ Ð½Ð° Ð¸Ð¼Ð¼ÑÐ½Ð¾Ð°ÑÑÐ¸Ð½Ð½Ð¾Ð¼ Ð·Ð°ÑÐ²Ð°ÑÐµ. Ð¡Ð¾Ð´ÐµÑÐ¶Ð°Ð½Ð¸Ðµ Ð¼Ð¸ÐºÑÐ¾Ð ÐÐ Ð² ÑÐ¿ÐµÑÐ¸ÑÐ¸ÑÐµÑÐºÐ¸Ñ ÑÑÐ°ÐºÑÐ¸ÑÑ Ð²ÐµÐ·Ð¸ÐºÑÐ» Ð¾ÑÐµÐ½Ð¸Ð²Ð°Ð»Ð¸ Ð¼ÐµÑÐ¾Ð´Ð¾Ð¼ ÐÐ¢-ÐÐ¦Ð . ÐÐ±ÑÐ°Ð±Ð¾ÑÐºÐ° Ð¸ Ð°Ð½Ð°Ð»Ð¸Ð· Ð´Ð°Ð½Ð½ÑÑ Ð¾ÑÑÑÐµÑÑÐ²Ð»ÑÐ»Ð¸ÑÑ Ð² Ð¿ÑÐ¾Ð³ÑÐ°Ð¼Ð¼Ð°Ñ NTA 2.3 Software, CytExpert, ImageJ, CFX Manager Software 3.1, SigmaPlot 12.0 Ð¸ OriginPro 9.1. Ð ÑÐµÐ·ÑÐ»ÑÑÐ°ÑÐµ, Ð±ÑÐ»Ð¸ Ð¾Ð¿ÑÐµÐ´ÐµÐ»ÐµÐ½Ñ ÑÐ¸Ð·Ð¸ÑÐµÑÐºÐ¸Ðµ Ð¸ Ð±Ð¸Ð¾ÑÐ¸Ð¼Ð¸ÑÐµÑÐºÐ¸Ðµ ÑÐ°ÑÐ°ÐºÑÐµÑÐ¸ÑÑÐ¸ÐºÐ¸ ÑÐ¾ÑÐ°Ð»ÑÐ½Ð¾Ð¹ Ð¿Ð¾Ð¿ÑÐ»ÑÑÐ¸Ð¸ Ð²ÐµÐ·Ð¸ÐºÑÐ». Ð¢ÐÐ(+) ÑÑÐ°ÐºÑÐ¸Ñ ÐÐÐ Ð¿Ð¾Ð»ÑÑÐµÐ½Ð° Ð¼ÐµÑÐ¾Ð´Ð¾Ð¼ Ð¼Ð°Ð³Ð½Ð¸ÑÐ¾Ð¸Ð¼Ð¼ÑÐ½Ð¾Ð¿ÑÐµÑÐ¸Ð¿Ð¸ÑÐ°ÑÐ¸Ð¸. Ð Ð°Ð·ÑÐ°Ð±Ð¾ÑÐ°Ð½Ð° Ð½Ð¾Ð²Ð°Ñ Ð¼ÐµÑÐ¾Ð´Ð¸ÐºÐ° Ð´Ð»Ñ Ð²ÑÐ´ÐµÐ»ÐµÐ½Ð¸Ñ ÐÐ¡ÐÐ(+) ÐÐÐ Ð½Ð° Ð¾ÑÐ½Ð¾Ð²Ðµ ÐÐÐ-Ð°Ð¿ÑÐ°Ð¼ÐµÑÐ° Ð¸ Ð¼Ð°Ð³Ð½Ð¸ÑÐ½ÑÑ ÑÐ°ÑÑÐ¸Ñ, Ð¸ÑÐ¿Ð¾Ð»ÑÐ·ÑÑ ÐºÐ¾ÑÐ¾ÑÑÑ Ð¿Ð¾Ð»ÑÑÐ¸Ð»Ð¸ ÐÐ¡ÐÐ(+) ÑÑÐ°ÐºÑÐ¸Ñ ÐÐÐ. ÐÐ½Ð°Ð»Ð¸Ð· Ð¿Ð¾ÑÐµÐ½ÑÐ¸Ð°Ð»ÑÐ½Ð¾ Ð¼Ð°ÑÐºÐµÑÐ½ÑÑ Ð¼Ð¸ÐºÑÐ¾Ð ÐÐ, Ð² ÑÐ¿ÐµÑÐ¸ÑÐ¸ÑÐµÑÐºÐ¸Ñ ÑÑÐ°ÐºÑÐ¸ÑÑ Ð½Ð°Ð½Ð¾Ð²ÐµÐ·Ð¸ÐºÑÐ» Ð¾ÐºÐ°Ð·Ð°Ð»ÑÑ Ð±Ð¾Ð»ÐµÐµ ÑÑÑÐµÐºÑÐ¸Ð²ÐµÐ½, ÑÐµÐ¼ Ð² Ð¾Ð±ÑÐµÐ¹ Ð¿Ð¾Ð¿ÑÐ»ÑÑÐ¸Ð¸, ÑÑÐ¾ Ð¼Ð¾Ð¶ÐµÑ Ð¿Ð¾ÑÐ»ÑÐ¶Ð¸ÑÑ Ð¾ÑÐ½Ð¾Ð²Ð¾Ð¹ Ð´Ð»Ñ ÑÐ¾Ð·Ð´Ð°Ð½Ð¸Ñ Ð½Ð¾Ð²Ð¾Ð¹ ÑÑÑÐ°ÑÐµÐ³Ð¸Ð¸ Ð°Ð½Ð°Ð»Ð¸Ð·Ð° ÐÐÐ Ð¿Ð»Ð°Ð·Ð¼Ñ Ð¸ ÑÐ°Ð·ÑÐ°Ð±Ð¾ÑÐºÐ¸ Ð½Ð¾Ð²ÑÑ Ð¼ÐµÑÐ¾Ð´Ð¾Ð² Ð½ÐµÐ¸Ð½Ð²Ð°Ð·Ð¸Ð²Ð½Ð¾Ð¹ Ð´Ð¸Ð°Ð³Ð½Ð¾ÑÑÐ¸ÐºÐ¸ Ð¾Ð½ÐºÐ¾Ð»Ð¾Ð³Ð¸ÑÐµÑÐºÐ¸Ñ Ð·Ð°Ð±Ð¾Ð»ÐµÐ²Ð°Ð½Ð¸Ð¹.
description: This work is devoted to the study of the prospects for using microRNAs from specific fractions of extracellular nanovisicles (EVs) of plasma as markers for the dif-ferential diagnosis of various types of cancer. The tasks that were solved in the course of the study: 1. Isolation and analysis of the general population of nanovesicles. 2. Development of a method for the isolation of specific fractions of nanovesi-cles. 3. Isolation of TPO (+) and PSMA (+) specific fractions of nanovesicles. 4. Assessment of the diagnostic potential of using microRNAs from TPO (+) and PSMA (+) fractions of nanovisicles. The work was carried out on the basis of the NMRC of Oncology named after N.N. Petrov. The total population of vesicles was obtained by ultracentrifugation, two-phase polymer system, and size exclusion chromatography. Specific vesicle frac-tions were isolated using immunoaffinity uptake techniques. The content of microRNA in specific fractions of vesicles was assessed by RT-PCR. Data processing and analysis were carried out using NTA 2.3 Software, CytExpert, ImageJ, CFX Manager Software 3.1, SigmaPlot 12.0, and OriginPro 9.1 programs. As a result, the physical and biochemical characteristics of the total vesicle pop-ulation were determined. TPO (+) fraction EVs was obtained by the method of mag-netoimmunoprecipitation. A new method was developed for the isolation of PSMA (+) EVs based on a DNA aptamer and magnetic particles, using which the PSMA (+) fraction of EVs was obtained. The analysis of potentially marker microRNAs in spe-cific fractions of nanoviesicles proved to be more efficient than in the total population, that provides a platform for a new strategy of ENV analysis and new methods for non-invasive cancer diagnostic.