Examinando por Materia "Caco-2"

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  • Miniatura
    PublicaciónAcceso abierto
    Formation of three-dimensional cell culture of colorectal carcinoma cells for possible applications in alternative cancer treatment in vitro
    (Universidad EIA, 2023) Parias Carreño, Andrés Felipe; Toro, Lenka
    ABSTRACT: according to the Global Cancer Observatory, colorectal cancer (CRC) is the third most recurrent cancer worldwide and in Colombia. For 2022, the American Cancer Society previewed that it will be the cause of death of 52,580 persons. Current conventional treatments can generate adverse effects and there is a need for new alternative methods to fight this disease. In order to test these new alternative methods, there is a need for a model that correctly represents the human tissue microenvironment to determine the possible effect of these new treatments. 3D models have the ability to form cell-to-cell interactions, mimic in vivo inflammatory response, develop drug resistance and allows the possibility for evaluating the treatment targeting capabilities. In this work the use of 3D cancer cell cultures is proposed as a possible model for the evaluation of novel alternative methods to fight colorectal cancer. The main objective of this project is to evaluate the ability of cancer cell line Caco-2 to form a 3D cell culture for possible applications in alternative colorectal cancer treatment. For the project execution, the culture conditions for 2D cell cultures were established based on a literature review in order to determine the 3D cell culture conditions. For the formation of the 2D culture and subcultures different experimental setups were tested to identify the best conditions for the 2D cell culture. The 3D cell culture was formed using the hanging drops method in droplets of 25 µL to obtain Caco-2 derived spheroids. The spheroids growth formed with different cell concentrations was observed to determine which conditions provided the best results for the formation of the spheroids. After the spheroids were formed the viability was evaluated by measuring the metabolic activity of the cells in the spheroids and the response to fluorescent dyes. To evaluate the proliferation a measure of the diameter of a single cell inside the droplet was recorded in day 1 and compared to the diameter of the spheroids formed. Morphology was observed under Scanning Electron Microscopy and all the results were evaluated using statistical analysis to determine the significance. As a result, Caco-2 derived spheroids were formed in 6 days under different cell concentrations with average diameters ranged between 70 and 80 µm in complete culture media supplemented with 20% FBS and 0.1% gentamycin with no additional supplements.
  • Miniatura
    PublicaciónAcceso abierto
    Protocol for the evaluation of chitosan nanoparticles in colorectal cancer cells
    (Universidad EIA, 2023) Avendaño Rodriguez, Jose Miguel; Flórez Ibáñez, María Paz; Echeverri Cuartas, Claudia Elena; Toro, Lenka
    ABSTRACT: Colorectal cancer is the third most frequent type of cancer in the world and in Colombia. According to the World Health Organization (WHO), in 2021 approximately 2 million people were reported with this condition, and, in Colombia, the figure rises to 2,000 new cases per year. Despite being a disease with high prevalence, the treatments that have been developed to combat it are not very effective, generating the need to explore new alternatives, such as nanotechnology that has allowed advances, however, the search for materials and techniques that allow an effective performance in the treatment of colorectal cancer continues. In this work the use of chitosan nanoparticles was proposed because they have advantages in stability, biodegradability, and biocompatibility, besides they can be crossed with other molecules to improve their properties. Therefore, the objective of this project was to establish a protocol to evaluate chitosan-based nanoparticles in a culture of colorectal cancer cells for their possible application as a treatment for this disease. To carry out the project, a methodology divided into two stages was proposed, the first stage consisted of obtaining and analyzing the stability of the nanoparticles, and the second stage consisted of evaluating the nanoparticles in two cell line cultures (Caco-2 cancer cell line and the NIH-3T3 mouse fibroblast cell line). The evaluation was performed at different concentrations of nanoparticles in a range between 0 - 100 µg/mL to establish the experimental conditions for the study of the effect of these nanoparticles on the cell lines mentioned, and thus contribute to the development of the project entitled "Chitosan - Fe (III) nanocomplexes modified with folic acid for targeted therapy in colon cancer" of the research group GIBEC of the EIA University. The results obtained showed that the ionotropic gelation method used to synthesize chitosan nanoparticles allows obtaining particles with an average size of 188.3 nm. However, when in contact with the protein-free cell culture medium, an increase in size was observed, reaching approximately 902.2 nm, with an average ζ-potential value of 10.79 mV. Furthermore, upon evaluating these nanoparticles in the cell cultures mentioned previously, it was possible to verify that they do not exert cytotoxicity on the cell lines studied, since in all the tests performed, the cell viability results were never lower than 70%.