Estandarización de un protocolo basado en el análisis de la proliferación de linfocitos t como modelo de estudio para la selección de principios activos con actividad inmunomoduladora

Trujillo Vargas, Angie Patricia

Estandarización de un protocolo basado en el análisis de la proliferación de linfocitos t como modelo de estudio para la selección de principios activos con actividad inmunomoduladora

dc.contributor.advisor	Morantes Medina, Sandra Johanna
dc.contributor.advisor	Delgado Tiria, Félix Giovanni
dc.contributor.author	Trujillo Vargas, Angie Patricia
dc.date.accessioned	2025-05-16T17:31:47Z
dc.date.available	2025-05-16T17:31:47Z
dc.date.issued	2025-05
dc.description.abstract	Los linfocitos T juegan un papel fundamental en la defensa del cuerpo al identificar y eliminar sustancias extrañas, además de regular la activación de otras células del sistema inmune para evitar respuestas patológicas. En este sentido, es esencial contar con protocolos que midan su actividad, a través de la estimulación o inhibiendo la proliferación de los Linfocitos T. El ensayo con carboxifluoresceína succinimidil éster(CFSE) se utiliza ampliamente para este fin, aunque con diversas variaciones en su implementación. En la presente investigación, se definieron los parámetros experimentales para evaluar la proliferación de linfocitos T en un modelo de reacción mixta leucocitaria utilizando CFSE. Variables como la concentración del reactivo, tiempo de incubación y proporción de células dendríticas (CD) y linfocitos T (LT) fueron analizados. Inicialmente, se establecieron las condiciones experimentales para la adecuada incorporación de CFSE a una concentración de 1,5μM, 2,5μM y 5μM, durante un período de 10, 15 y 20 minutos. La proliferación fue monitoreada cada 48 horas mediante citometría de flujo, siguiendo la reducción progresiva de la fluorescencia. . Se emplearon diferentes proporciones entre células dendríticas (CD) y linfocitos T (LT) (CD:LT de 2:1, 1:1 y 1:2), efectuando un seguimiento periódico por citometría de flujo durante 8 días. Finalmente se evidenció que los LT incorporaron el reactivo CFSE a una concentración de 5μM y un tiempo de incubación de 10 min. Finalmente se demostró, que tanto la proporción como el tiempo influyen de manera significativa en el seguimiento de la proliferación de los linfocitos T en un cultivo de reacción mixta leucocitaria. En consecuencia, se recomienda emplear una proporción CD:LT de de 2:1, 5μM de CFSE y 10 min de incubación en futuras investigaciones
dc.description.abstractenglish	T lymphocytes play a fundamental role in the body's defense by identifying and eliminating foreign substances, as well as regulating the activation of other cells of the immune system to avoid pathological responses. In this sense, it is essential to have protocols that measure their activity by stimulating or inhibiting the proliferation of T lymphocytes. The carboxyfluorescein succinimidyl ester (CFSE) assay is widely used for this purpose, although with several variations in its implementation. In the present investigation, experimental parameters were defined to assess T lymphocyte proliferation in a mixed leukocyte reaction model using CFSE. Variables such as reagent concentration, incubation time and proportion of dendritic cells (DC) and T lymphocytes (LT) were analyzed. Initially, experimental conditions were established for proper incorporation of CFSE at a concentration of 1.5μM, 2.5μM and 5μM, for a period of 10, 15 and 20 minutes. Proliferation was monitored every 48 hours by flow cytometry, following the progressive reduction of fluorescence. Different ratios of dendritic cells (DC) to T lymphocytes (LT) were used (DC:LT of 2:1, 1:1 and 1:2), with periodic monitoring by flow cytometry for 8 days. Finally, it was evidenced that LTs incorporated the CFSE reagent at a concentration of 5μM and an incubation time of 10 min. Finally, it was shown that both ratio and time significantly influence the monitoring of T lymphocyte proliferation in a mixed leukocyte reaction culture. Consequently, it is recommended to use a CD:LT ratio of 2:1, 5μM of CFSE and 10 min incubation in future investigations.
dc.description.degreelevel	Pregrado	spa
dc.description.degreelevel	Químico Farmacéutico	spa
dc.format.mimetype	application/pdf
dc.identifier.instname	Universidad El Bosque	spa
dc.identifier.reponame	reponame:Repositorio Institucional Universidad El Bosque	spa
dc.identifier.repourl	repourl:https://repositorio.unbosque.edu.co
dc.identifier.uri	https://hdl.handle.net/20.500.12495/14372
dc.language.iso	es
dc.publisher.faculty	Facultad de Ciencias	spa
dc.publisher.grantor	Universidad El Bosque	spa
dc.publisher.program	Química Farmacéutica	spa
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dc.rights	Attribution 4.0 International	en
dc.rights.accessrights	info:eu-repo/semantics/openAccess
dc.rights.accessrights	https://purl.org/coar/access_right/c_abf2
dc.rights.local	Acceso abierto	spa
dc.rights.uri	http://creativecommons.org/licenses/by/4.0/
dc.subject	Linfocitos T
dc.subject	Reactivo CFSE
dc.subject	Células dendríticas
dc.subject	Reacción mixta leucocitaria
dc.subject	Fluorescencia
dc.subject.ddc	615.19
dc.subject.keywords	T lymphocytes
dc.subject.keywords	CFSE Reagent
dc.subject.keywords	Dendritic cells
dc.subject.keywords	Mixed leukocyte reaction
dc.subject.keywords	Fluorescence
dc.title	Estandarización de un protocolo basado en el análisis de la proliferación de linfocitos t como modelo de estudio para la selección de principios activos con actividad inmunomoduladora
dc.title.translated	Standardization of a protocol based on the analysis of t lymphocyte proliferation as a study model for the selection of active ingredients with immunomodulatory activity
dc.type.coar	https://purl.org/coar/resource_type/c_7a1f
dc.type.coarversion	https://purl.org/coar/version/c_ab4af688f83e57aa
dc.type.driver	info:eu-repo/semantics/bachelorThesis
dc.type.hasversion	info:eu-repo/semantics/acceptedVersion
dc.type.local	Tesis/Trabajo de grado - Monografía - Pregrado