Exosome Isolation and RNA Extraction

Exosomes from human and mouse plasma were extracted using an Invitrogen™ Total Exosome Isolation Kit (ThermoFisher, USA) [27 (link)]. Briefly, 200 uL of plasma was mixed with the precipitation reagent provided by the kit and exosomes were isolated according to the manufacturer’s instructions. The exoRNeasy Serum/Plasma Maxi Kit (Yeasen Biotech, Shanghai, China) [28 (link)] was employed to isolate exosomal RNAs in samples previously spiked with 25 fmol of C. elegans cel-miR-39 standard RNA (Sangon, Shanghai, China), used as control. The exosomes were stored at -80° C before cell culture experiments and molecular analyses.

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Cao W., Zeng Z., He Z, & Lei S. (2021). Hypoxic pancreatic stellate cell-derived exosomal mirnas promote proliferation and invasion of pancreatic cancer through the PTEN/AKT pathway. Aging (Albany NY), 13(5), 7120-7132.

Publication 2021

Invitrogen™ Total Exosome Isolation Kit exoRNeasy Serum/Plasma Maxi Kit C. elegans cel-miR-39 standard RNA

Cell culture Exosomes Human Isolation Mouse Plasma Serum

Corresponding Organization :

Other organizations : Guiyang Medical University, Affiliated Hospital of Guizhou Medical University

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Variable analysis

independent variables

Exosome extraction method using Invitrogen™ Total Exosome Isolation Kit
Exosomal RNA isolation using exoRNeasy Serum/Plasma Maxi Kit

dependent variables

Exosome extraction
Exosomal RNA isolation

control variables

Plasma sample volume (200 uL)
Spike-in of 25 fmol of C. elegans cel-miR-39 standard RNA

controls

Positive control: C. elegans cel-miR-39 standard RNA
Negative control: Not explicitly mentioned

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