Ciencias Exactas y Ciencias de la Salud
Permanent URI for this collectionhttps://hdl.handle.net/11285/551039
Pertenecen a esta colección Tesis y Trabajos de grado de las Maestrías correspondientes a las Escuelas de Ingeniería y Ciencias así como a Medicina y Ciencias de la Salud.
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- Recovery and purification of exosomes using polymer-polymer aqueous two-phase systems (ATPS)(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2020-12-03) Torres Bautista, Abril Lorena; González Valdez, José Guillermo; emipsanchez/tolmquevedo; Torres Acosta, Mario Antonio; Mejía Manzano, Luis Alberto; Ramos de la Peña, Ana Mayela; Escuela de Ingeniería y Ciencias; Campus MonterreyExosomes are a type of extracellular vesicles that present natural and attractive characteristics such as nano size (30-200 nm) and unique structure for their use as drug delivery systems for drug therapy, biomarkers for prognostic, diagnostic and personalized treatments. So far, one of the major challenges for therapeutic applications of exosomes is the development of optimized methods for their isolation. In this context, Aqueous Two-Phase Systems (ATPS) have shown to be an alternative method to isolate biological molecules and particles with promising expectations for exosomes. In this work, fractionation of exosomes obtained from a CaCo2 cell line and the contaminants present in their culture media was performed in 20 different polymer-polymer aqueous two-phase systems (ATPSs) in a discrete way. The effect of system design parameters such as polymer composition, molecular weight, and TLL (15-30%) on PEG-Dextran, Dextran-Ficoll and PEG-Ficoll systems were studied keeping phase volume ratio (VR) and pH constant at 1.0 and 7.0, respectively. After partition analysis, 4 out of the 20 systems presented the best exosome fractionation from the contaminants under initial conditions and were selected for further partition optimization via salt addition (NaCl) to observe the behavior of both samples, individually, and improve the efficiency of collection. The PEG 10,000 gmol-¹ – Dextran 10,000 gmol-¹ system at TLL 25% w/w, with the addition NaCl to reach a final concentration of 25 mM, showed the best isolation efficiency resulting in exosome purification factor of 2 and furtherly demonstrated that ATPS has the potential for the selective recovery these promising nanovesicles.
- Exosome-mediated insulin delivery for the treatment of diabetes mellitus(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2020-11-16) Rodríguez Morales, Belén; GONZALEZ VALDEZ, JOSE GUILLERMO; 234501; González Valdez, José Guillermo; tolmquevedo, emipsanchez; Hernández Pérez, Jesús; Rodríguez Sánchez, Dariana Graciela; School of Engineering and Sciences; Campus Monterrey; Antunes Ricardo, MarilenaExosomes are membrane-bound extracellular nanovesicles of 30-150 nm released by almost all cell types. Since they were discovered in 1983, numerous studies yielded clear evidences that exosomes serve as essential messengers for hundreds of biological-signaling processes as well as in pathological processes. In consequence of the discovery of exosomal natural functions, a wide range of applications have been developed in several fields of study, especially in drug delivery for therapeutic use. Following this growing stream of study, the aim of this work was to test the efficiency of exosome-mediated human insulin delivery using exosomes extracted from three different cell lines: hepatocellular carcinoma (HepG2); primary dermal fibroblasts (HDFa) and pancreatic β cells (RIN-m); all related to the production and/or the ability to sense insulin and, consequently regulate glucose levels in the extracellular medium. The obtained results revealed that the optimal loading efficiency was achieved by a 200 V electroporation, in comparison to an incubation at room temperature. Moreover, the maximum in vitro exosome uptake was reached after 6 h of incubation with loaded exosomes, and lightly decrease 24 h after adding the exosomes. Furthermore, glucose quantification assays revealed that exosome-mediated incorporation of insulin present significative differences in HDFa and HepG2 cells, enhancing the transport in HDFa, in comparison with free human insulin effect in the regulation of extracellular glucose levels. On the other hand, no significative differences were found between both treatments in RIN-cells. Hence, the results suggest that exosomes could be a potentially valuable tool for a biocompatible and stable insulin delivery as part of a future treatment of diabetes mellitus.