185 Overexpression or CRISPr/Cas9-mediated inhibition of prostaglandin E2 receptors EP2 and EP4 in equine adipose mesenchymal stem cells: implications for cell migration and cellular therapies
A. C. F. Mançanares A , J. O. Manríquez A , J. Cabezas A , F. Telleria B , L. Rodriguez A and F. O. Castro AA Animal Science, University of Concepcion, Chillán, Chile;
B Ingeneria en Biotecnologia, Facultad de Ciencias Agrarias y Forestales, Universidad Catolica del Maule, Región del Maule, Talca, Chile
Reproduction, Fertility and Development 31(1) 217-217 https://doi.org/10.1071/RDv31n1Ab185
Published online: 3 December 2018
Abstract
Prostaglandin E2 (PGE2) acts through 4 cellular receptors: EP1, EP2, EP3, and EP4; only EP2 and EP4 are relevant for immunomodulation and migration in immune cells. Besides those, several cells express these receptors on their surface, including mesenchymal stem cells. Pharmacological inhibition of the EP2 receptor prevents migration of immune system cells to inflamed sites, where the concentration of PGE2 is high. Based on this, we hypothesised that overexpression of EP2 or EP4 receptors in equine mesenchymal stem cells (eMSC) will improve their migration to inflammatory sites and subsequent homing capability. Conversely, their suppression will lead to low or no migration, favouring the paracrine properties of MSC in the processes of tissue regeneration and reduction of inflammation. To test this, we manipulated the PGE2-EP2-EP4 axis and evaluated the effect of such modifications on transgenic cells in vitro. Equine MSC from adipose tissue were obtained from 5 animals. The coding sequences of both receptors were synthesised (GenScript, Hong Kong) based on the published horse genome (National Center for Biotechnology Information;
This research was supported by FONDECYT 3170390 to ACFM, Ministry of Education, Chile.