Publicação
Effect of mesenchymal stem cell source on the molecular profile of extracellular vesicles for cell therapy
| Resumo: | Mesenchymal stem cells (MSCs) obtained from different sources have been considered for cell therapeutics against several diseases, were its derived extracellular vesicles (EVs) are the primary mediators of the cells paracrine effect. The present work aimed to evaluate the influence of different MSC sources, as adipose tissue, bone marrow and umbilical cord matrix on the molecular profile of EVs acquired by Fourier-Transform Infrared (FTIR) Spectroscopy after spectra pre-processing and its principal component and hierarchical cluster analysis. It was evaluated the EVs, MSCs and conditioned media molecular profiles in function of MSC source, MSC donor (i.e. 3 donors per source) and media composition (xenogeneic-free (XF) vs serum-containing medium (DMEM-FBS)). Two replicates cultures per donor were considered. EVs were obtained from the MSC conditioned media based on a kit assay using a polymer that, however, absorbs on the EVs surface leading to an interference on the EVs spectra signal. The EVs high dilution factor leads to the polymer partial desorption, as observed by other authors. Due to variability of the desorption process, a lower reproductible spectra from EVs were obtained. Despite that, it was possible to observe that EVs molecular profile depended on the media type and, for each MSC source, on the MSC donor. It was also observed that EVs molecular profile obtained from DMEM cultures apparently presented a more homogenous chemical profile than EVs obtained with XF medium. Diverse ratios between infrared spectral bands were identified, that discriminate at a 5% significance, in mean terms, the EVs molecular profile in function of the culture medium composition. The MSC spectra were also highly variable due to different growth patterns. Even so, it was possible to observe that MSC molecular profile depended on the culture media type, being observed a more homogeneous molecular profile when cells were cultured in XF than in DMEM medium. When grown on DMEM medium, it was possible to observe that MSC molecular profile depended on the MSC source. MSC molecular profile also apparently depended on the MSC donor. A very similar conditioned media spectra were obtained from the different cultures and, consequently, higher reproducible molecular profiles were obtained in relation to MSC and EVs analyses. It was observed that conditioned media molecular profile depended on MSC source, MSC donor and culture media composition. A lower variable molecular profile was observed when using XF medium, as observed with MSC molecular profile. From the EVs, MSC and conditioned media molecular profiles, the best reproductive results were obtained from the conditioned media, that pointed that it was affected by the MSC donor and source. The impact of the culture media type, MSC source and donor was also partially observed on the EVs and MSC molecular profile. All these results point to the impact of the culture media, MSC source and donor on the EVs molecular profile. Therefore, in therapies-cell based and based EVs-based, the impact of culture media, MSC source and donor should be considered. |
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| Autores principais: | Pires, Filipa Adriana Rego |
| Assunto: | Mesenchymal stem cells Células estaminais mesenquimais Extracellular vesicles Vesículas extracelulares FTIR Spectroscopy Espectroscopia de IVT Molecular profile Perfil molecular, |
| Ano: | 2019 |
| País: | Portugal |
| Tipo de documento: | dissertação de mestrado |
| Tipo de acesso: | acesso aberto |
| Instituição associada: | Instituto Politécnico de Lisboa |
| Idioma: | inglês |
| Origem: | Repositório Científico do Instituto Politécnico de Lisboa |
| Resumo: | Mesenchymal stem cells (MSCs) obtained from different sources have been considered for cell therapeutics against several diseases, were its derived extracellular vesicles (EVs) are the primary mediators of the cells paracrine effect. The present work aimed to evaluate the influence of different MSC sources, as adipose tissue, bone marrow and umbilical cord matrix on the molecular profile of EVs acquired by Fourier-Transform Infrared (FTIR) Spectroscopy after spectra pre-processing and its principal component and hierarchical cluster analysis. It was evaluated the EVs, MSCs and conditioned media molecular profiles in function of MSC source, MSC donor (i.e. 3 donors per source) and media composition (xenogeneic-free (XF) vs serum-containing medium (DMEM-FBS)). Two replicates cultures per donor were considered. EVs were obtained from the MSC conditioned media based on a kit assay using a polymer that, however, absorbs on the EVs surface leading to an interference on the EVs spectra signal. The EVs high dilution factor leads to the polymer partial desorption, as observed by other authors. Due to variability of the desorption process, a lower reproductible spectra from EVs were obtained. Despite that, it was possible to observe that EVs molecular profile depended on the media type and, for each MSC source, on the MSC donor. It was also observed that EVs molecular profile obtained from DMEM cultures apparently presented a more homogenous chemical profile than EVs obtained with XF medium. Diverse ratios between infrared spectral bands were identified, that discriminate at a 5% significance, in mean terms, the EVs molecular profile in function of the culture medium composition. The MSC spectra were also highly variable due to different growth patterns. Even so, it was possible to observe that MSC molecular profile depended on the culture media type, being observed a more homogeneous molecular profile when cells were cultured in XF than in DMEM medium. When grown on DMEM medium, it was possible to observe that MSC molecular profile depended on the MSC source. MSC molecular profile also apparently depended on the MSC donor. A very similar conditioned media spectra were obtained from the different cultures and, consequently, higher reproducible molecular profiles were obtained in relation to MSC and EVs analyses. It was observed that conditioned media molecular profile depended on MSC source, MSC donor and culture media composition. A lower variable molecular profile was observed when using XF medium, as observed with MSC molecular profile. From the EVs, MSC and conditioned media molecular profiles, the best reproductive results were obtained from the conditioned media, that pointed that it was affected by the MSC donor and source. The impact of the culture media type, MSC source and donor was also partially observed on the EVs and MSC molecular profile. All these results point to the impact of the culture media, MSC source and donor on the EVs molecular profile. Therefore, in therapies-cell based and based EVs-based, the impact of culture media, MSC source and donor should be considered. |
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