Publicação
The role of exosomes in cellular communication
| Resumo: | Exosomes are extracellular vesicles responsible for transporting biological cargo between cells and may be present in all body fluids. As such, exosomes have been indicated in many bodily functions as well as in pathological roles. This makes them a particularly exciting target for research, as learning more about them can permit the formulation of new treatments, diagnostic methods, or drug-delivery systems with exosomes as nanocarriers. These vesicles have been previously associated with various stages of cancer progression, including the establishment of the pre-metastatic niche and tumor homing. Here, the work focused on evaluating the possibility of cancer cells inducing an exosome-mediated preparation of the pre-metastatic niche using a proxy (intermediary courier) healthy cell (Fibroblasts). A secondary aim of the study was to assess the potential of exosomes as nanocarriers for gold nanoconjugates carrying therapeutic moieties. Here, we validated a protocol of exosome extraction method that uses a combination of Size-Based methods and a Precipitating-Based commercial kit. We did so by characterizing the resultant exosomes using various techniques, including Dynamic Light Scattering, Western-Blot, and two commercial exosome quantification kits. In the study of the metastatic role of exosomes, the results are inconsistent with what was to be expected, suggesting a lower C-MYC expression after incubation with malignant exosomes. However, this is probably the reflection of experimental errors and thus, further repetitions of this experiment are needed. In the assessment of exosomes as nanocarriers, we successfully achieved encapsulation of gold nanoparticles inside exosomes, as visualized by Dark-Field Fluorescence Microscopy, but not with the intended efficiency. Further optimization of the incubation periods and process is required. Due to the CoVID-19 pandemic, which caused severe constraints to the intensity of laboratory work, the full plan could not be implemented to completion. However, several relevant cues were provided to allow future work within this line of reasoning. |
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| Autores principais: | Costa, João Pedro Gonzalez León Lapa e |
| Assunto: | Exossomas Vesículas extracelulares Metástase Nanopartículas Teses de mestrado - 2020 |
| Ano: | 2021 |
| País: | Portugal |
| Tipo de documento: | dissertação de mestrado |
| Tipo de acesso: | acesso aberto |
| Instituição associada: | Universidade de Lisboa |
| Idioma: | inglês |
| Origem: | Repositório da Universidade de Lisboa |
| Resumo: | Exosomes are extracellular vesicles responsible for transporting biological cargo between cells and may be present in all body fluids. As such, exosomes have been indicated in many bodily functions as well as in pathological roles. This makes them a particularly exciting target for research, as learning more about them can permit the formulation of new treatments, diagnostic methods, or drug-delivery systems with exosomes as nanocarriers. These vesicles have been previously associated with various stages of cancer progression, including the establishment of the pre-metastatic niche and tumor homing. Here, the work focused on evaluating the possibility of cancer cells inducing an exosome-mediated preparation of the pre-metastatic niche using a proxy (intermediary courier) healthy cell (Fibroblasts). A secondary aim of the study was to assess the potential of exosomes as nanocarriers for gold nanoconjugates carrying therapeutic moieties. Here, we validated a protocol of exosome extraction method that uses a combination of Size-Based methods and a Precipitating-Based commercial kit. We did so by characterizing the resultant exosomes using various techniques, including Dynamic Light Scattering, Western-Blot, and two commercial exosome quantification kits. In the study of the metastatic role of exosomes, the results are inconsistent with what was to be expected, suggesting a lower C-MYC expression after incubation with malignant exosomes. However, this is probably the reflection of experimental errors and thus, further repetitions of this experiment are needed. In the assessment of exosomes as nanocarriers, we successfully achieved encapsulation of gold nanoparticles inside exosomes, as visualized by Dark-Field Fluorescence Microscopy, but not with the intended efficiency. Further optimization of the incubation periods and process is required. Due to the CoVID-19 pandemic, which caused severe constraints to the intensity of laboratory work, the full plan could not be implemented to completion. However, several relevant cues were provided to allow future work within this line of reasoning. |
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