CANTERO MarÍa JosÉ
congresos y reuniones científicas
MESENCHYMAL STROMAL CELLS AS CELLULAR CARRIERS OF THERAPEUTIC GENES: USE OF IN VITRO TRANSCRIBED mRNA
LLAMAZARES GONZALEZ LUCRECIA; CANTERO MARIA JOSE; FIORE ESTEBAN; BAYO JUAN; DOMINGUEZ LUCIANA; ATORRASAGASTI CATALINA; MALVICINI MARIANA; MAZZOLINI GUILLERMO; GARCIA MARIANA GABRIELA
Mar del Plata
Congreso; Reunión Conjunta de SAIC SAI SAFIS 2018; 2017
The use of mesenchymal stromal cells (MSCs) has been proposed during the last years for therapeutic purposes, especially for regenerative therapies. They are promising candidates as vehicles for delivery of therapeutic agents. The majority of engineering strategies have focused on use of viral vector for gene transfer. However, the use of in vitro transcribed mRNA (IVT mRNA) is gaining attention as a promising tool for the delivery of therapeutic genes by MSCs. The aim of this work was to express the mRNA of insulin growth factor I (IGF-I) and DsRed in MSCs, with the aim to treat liver fibrosis. For this purpose, mRNA (of IGF-I or DsRed) was in vitro transcribed, modified to be more stable within the cell, and then transfected into human umbilical cord perivascular cells-derived MSCs with lipofectamine. IGF-I production by MSCs was evaluated by ELISA, and DsRed expression by fluorescence microscopy. Different quantities of mRNA were assayed, and we observed that low amount of mRNA (0.2 µg mRNA / 40.000 cells) were sufficient to express the exogenous proteins up to 15 days. Since the use of MSCs for therapeutic purposes relies on their capacity to migrate to the injured tissues, we also evaluated their migration capacity (using a modified Boyden chamber). We observed that MSCs expressing IGF-I or DsRed have the same migratory capacity compared with unmodified MSCs. Our results demonstrate that IVT mRNA is an efficient method to engineer human MSCs, and suggest that this approach would be useful for therapeutic purposes.