Microvesicles secreted from equine amniotic-derived cells and their potential role in reducing inflammation in endometrial cells in an in-vitro modelReport as inadecuate




Microvesicles secreted from equine amniotic-derived cells and their potential role in reducing inflammation in endometrial cells in an in-vitro model - Download this document for free, or read online. Document in PDF available to download.

Stem Cell Research and Therapy

, 7:169

First Online: 18 November 2016Received: 31 August 2016Revised: 22 September 2016Accepted: 21 October 2016

Abstract

BackgroundIt is known that a paracrine mechanism exists between mesenchymal stem cells and target cells. This process may involve microvesicles MVs as an integral component of cell-to-cell communication.

MethodsIn this context, this study aims to understand the efficacy of MVs in in-vitro endometrial stressed cells in view of potential healing in in-vivo studies. For this purpose, the presence and type of MVs secreted by amniotic mesenchymal stem cells AMCs were investigated and the response of endometrial cells to MVs was studied using a dose-response curve at different concentrations and times. Moreover, the ability of MVs to counteract the in vitro stress in endometrial cells induced by lipopolysaccharide was studied by measuring the rate of apoptosis and cell proliferation, the expression of some pro-inflammatory genes such as tumor necrosis factor-α TNF-α, interleukin-6 IL-6, interleukin 1β IL-1β, and metalloproteinases MMP 1 and 13, and the release of some pro- or anti-inflammatory cytokines.

ResultsMVs secreted by the AMCs ranged in size from 100 to 200 nm. The incorporation of MVs was gradual over time and peaked at 72 h. MVs reduced the apoptosis rate, increased cell proliferation values, downregulated pro-inflammatory gene expression, and decreased the secretion of pro-inflammatory cytokines.

ConclusionOur data suggest that some microRNAs could contribute to counteracting in-vivo inflammation of endometrial tissue.

KeywordsMicrovesicles Endometrium Inflammation LPS Regenerative medicine Abbreviationsa.u.Arbitrary units

AMCAmniotic mesenchymal stem cell

AOIArea of interest

CMConditioned medium

EDCEndometrial cell

EGFEpidermal growth factor

EXExosome

FBSFetal bovine serum

GAPDHGlyceraldehyde-3-phosphate dehydrogenase gene

HEPESN-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid

HG-DMEMHigh-glucose Dulbecco’s modified Eagle’s medium

Hoxa9Homeobox protein Hox-A9-like

ILInterleukin

LPSLipopolysaccharide

miRNAMicroRNA

MMPMatrix metalloproteinase

MPRMembrane-associated progesterone receptor

MSCMesenchymal stem cell

MTT3-4,5-dimethylthiazol- 2-yl-2,5-diphenyletrazolium bromide

MVMicrovesicle

PPassage

PBSPhosphate-buffered saline

PGRMC1Membrane-associated progesterone receptor

PRProgesterone Receptor

qRT-PCRQuantitative reverse transcription polymerase chain reaction

RT-PCRReverse transcription polymerase chain reaction

SVShedding vesicle

TEMTransmission electron microscopy

TGFTransforming growth factor

TLRToll-like receptor

TNFTumor necrosis factor

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Author: Claudia Perrini - Maria Giuseppina Strillacci - Alessandro Bagnato - Paola Esposti - Maria Giovanna Marini - Bruna Corrade

Source: https://link.springer.com/



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