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.
Browse
Search Results
- On the improvement of in vitro senescence studies by applying microfluidic technology and mathematical modelling(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2021-06-09) Rios Maciel, Mauricio Andrés; González Meljem, José Mario; puelquio; Álvarez, Mario Moisés; Trujillo de Santiago, Grissel; Escuela de Ingeniería y Ciencias; Campus MonterreyCellular senescence can be defined as an irreversible cell cycle arrest, which has been termed as an ‘antagonistic pleiotropy’ because of the duality of its role in aging. While the early-life on-set of the phenotype prevents tumorigenesis and plays an important part on the immune system-mediated removal of damaged cells, accumulation of senescent cells late in life is linked to the development of age-related diseases; thus, senescence is a target for anti-aging therapies. However, the senescent phenotype is very complex and heterogenous, making in vitro approaches desirable for the study of their individual components. In vitro culture of senescence presents a complex combination of different types of cells, including senescent, non-senescent, arrested and apoptotic, making it challenging to study the process in a complete manner. In addition, senescent cells have complex intercellular interactions, as well as ECM interactions. Recent approaches have used microfluidic technology alongside the incorporation of three-dimensional hydrogel-based matrixes to better mimic the microenvironment of the cell in vitro to make it more physiologically relevant. In this work we focus on the preliminary work for a further translation of traditional in vitro senescence techniques to a 3D and microfluidic environment. We assessed, analyzed and verified the behavior of endothelial HMEC-1 cells when cultured on GelMA-based hydrogel, as well as the effects of the matrix in senescence induction and assessment methods. We developed a mathematical model based on reported data to identify variables of importance and simulate the transition between states of proliferation, death and senescence through time in a senescent culture induced through doxorubicin. We observed that doxorubicin-induced senescence in endothelial cells stimulates apoptosis but yields an acceptable percentage of induction. After transferring these experiments to the GelMA-based hydrogel, neither induction nor assessment seemed to present a problem. We observed that endothelial cells in GelMA tend to adjust to a specific percentage, giving insight into the importance of the pore size in the matrix. Finally, the mathematical model shed light on the importance of time after treatment in the transition to senescent cells but also the turnover to dead population.
- Design of droplet-based microfluidic devices and its application for protein fractionation(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2020-12-04) Hernández Cid, David; Mata Gómez, Marco Arnulfo; qro /|bqrotbecerra/tolmquevedo; Pérez González, Victor Hugo; Cervantes Avilés, Pabel Antonio; School of Engineering and Science; Campus Monterrey; González Valdéz, José GuillermoMicrofluidics technology offers new possibilities due to the nature of small scale, such as the high surface area to volume ratio. Droplet based microfluidics is an emerging field that has found its application in different areas, like material science, chemical reactions, and biochemical analysis. On the other hand, downstream processing operations like separation and purification still has some drawbacks like big reagents consumption, huge waste generation, extra steps like concentration or salts removal. Here is where microfluidics can be implemented to overcome some of the issues presented in this field. In this work, we designed and developed a microfluidic platform able to generate droplet-like aqueous two- phases system (ATPS) in a continuous manner. We started this project by performing a study of how different forces like surface tension, inertial forces, viscous forces, and geometry affect droplet formation. We found that all the previously mentioned parameters have an impact in different ways on the process of droplet formation. Afterwards, this knowledge was used to implement a microfluidic flow-focusing on the generation of droplet-based micro ATPS with the used of salts and PEG as the two liquid phases of the ATPS. To see that this ATPS droplet microfluidic system really worked, we tested its capacity to fractionate ribonuclease A (RNase A) on its native and PEGylated forms, which have affinity for the salt and PEG phases, respectively. Thus, we expected that proteins would get inside or outside droplets or streamlines according to their affinity for each of the two phases. In doing so, native, and PEGylated proteins were labeled with FITC and their movement from one to another phase or their retention in the phase where protein was placed were recorded. It was confirmed that proteins move or keep in their phase for which they have more affinity. This result opens the possibility for the development of microfluidic separators based on ATPS droplet formation.
- Andamios porosos de zeína: prueba de concepto de su uso en el cultivo de células de mamífero.(2017-12-14) Portales Cabrera, Cynthia Guadalupe; Trujillo-de Santiago, Grissel.; Álvarez, Mario Moises; Montes Fonseca, Silvia Lorena; Mendoza Buenrostro, Christian CarlosLas espumas son materiales porosos y de baja densidad, características convenientes en andamiajes para células de mamífero. En este proyecto se estudió la aptitud de espumas elaboradas a partir de zeína (la proteína más abundante del maíz) como andamios para adherir y mantener distintos cultivos celulares de mamíferos: fibroblastos de ratón (NIH 3T3 ATCC® CRL-1658™), fibroblastos humanos FH (fuente primaria) y células de adenocarcinoma mamario MCF-7 (ATCC® HTB-22™). Los andamios porosos se obtuvieron a partir de zeína termoplastificada, la cual fue sometida a un proceso de expansión por dióxido de carbono supercrítico. Estos andamios fueron estudiados bajo condiciones de cultivo continuo en cámaras microfluídicas o lote en placas de ultrabaja adherencia. Los dispositivos microfluídicos, con un volumen efectivo de 100 µl, se fabricaron a partir de polidimetilsiloxano (PDMS) y se conectaron a una bomba de jeringa utilizando tubería flexible. Cortes rectangulares de espuma de zeína fueron colocados al interior de la cámara microfluídica, se inocularon con células de mamífero y se sometieron a condiciones de flujo continuo de medio modificado de Dulbecco (DMEM) a 3 µL/min. Resultados de microscopía confocal demuestran que todas las líneas celulares exploradas presentaron adhesión celular a los andamios de zeína. Resultados de consumo de glucosa confirman que las células mantuvieron su viabilidad en todos los experimentos, 60- 84 horas en cultivo continuo y hasta 9 días en cultivo por lote. Estos resultados sugieren que los andamios elaborados a base de zeína se pueden utilizar como un andamio adecuado para el cultivo de tejidos e ingeniería celular. Foams are low-density porous materials, a property that makes them attractive for their use as scaffolds for mammalian cells. In this project, micro foams made from thermo-plasticized zein (the main protein of maize) were used as scaffolds to attach and culture mouse fibroblasts (cell line NIH 3T3 ATCC® CRL-1658™), human fibroblasts (primary cell line FH), and adenocarcinoma cell line MCF-7 (ATCC® HTB-22™) in continuous flow chambers or ultra-low attachment multiwall plates (batch system). Zein foams were produced by supercritical-CO2 expansion. We produced polydimethylsiloxane (PDMS) devices with 100 µl of effective-volume micro-channels that were connected with tubing to a syringe pump to implement continuous culture experiments. Rectangular slices of zein foam were placed into the flow chamber, they were seeded with the corresponding cell line, and the devices were continuously perfused with Dulbecco's modified Eagle medium (DMEM) at a flow rate of 3 µl. The systems were incubated at 37°C and 5% CO2. Different behaviors were observed for each cell line, but cell adhesion was found in all experiments and it was documented using confocal microscopy. Trends of the glucose consumption curves show that the cultures maintained their viability throughout of experiment, 60-84 h in continuous cultures and up to 9 days in batch cultures. These proof-of-concept experiments suggest that zein can be used as a surface scaffold for continuous cell culture and tissue engineering.