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Effects of Different Centrifugal Forces on miRNA Components of Mouse Blastocysts-Derived Microvesicles
Liu Qing1, Wei Zixin1, Wang Yiting2, Ba Xiaoyun1, Li Hao1, Fang Liaoqiong1,2*
1College of Biotechnology, Southwest University, Chongqing 400716, China; 2National Engineering Research Center of Ultrasound Medicine, Chongqing 401121, China
Abstract: Microvesicle (MV) communication is associated with content composition. The microvesicles obtained by different preparation methods are different. The effects of different centrifugal forces on the size and miRNA components of MV from mouse blastocysts were analyzed. The MV of the blastocysts of d 3.5 was isolated and purified by ultracentrifugation at the force of 12 000 ×g， 50 000 ×g and 100 000 ×g respectively. The particle size range of MV was detected by particle size analyzer. The miRNA quantity of MV was analyzed by real-time PCR technique. The results showed that the average particle size of MV prepared by centrifugal force of 12 000 ×g was (444.5±258.7) nm， containing 333 miRNAs; 50 000 ×g of MV had an average particle size of (237.0±178.0) nm and contained 339 miRNAs; the average particle size of 100 000 ×g MV was (164.8±56.5) nm， 335 miRNAs. The size of the vesicles and the miRNA components are related to the centrifugal force. The miRNAs contained in the MVs obtained by different centrifugal forces and the abundance of the same miRNAs are different. Thus， different centrifugal forces affect the size of the prepared blastocysts MV and the components of the miRNA.
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