Avaliação do potencial de frações do soro humano em substituição ao soro fetal bovino no cultivo de células-tronco mesenquimais humanas

• Ano de defesa: 2019
• Autor(a) principal: Paiva, Bianca Simonassi Raso de
• Orientador(a): Pogue, Robert
• Tipo de documento: Dissertação
• Tipo de acesso: Acesso aberto
• Idioma: por
• Instituição de defesa: Universidade Católica de Brasília
• Programa de Pós-Graduação: Programa Stricto Sensu em Ciências Genômicas e Biotecnologia
• Departamento: Escola de Saúde e Medicina
• País: Brasil
• Palavras-chave em Português: Senescência; Soro humano; Células-tronco mesenquimais
• Palavras-chave em Inglês: Mesenchymal stem cell; Human serum; Senescence
• Área do conhecimento CNPq: CNPQ::CIENCIAS BIOLOGICAS::GENETICA
• Resumo em Inglês: Mesenchymal stem cells (MSCs) have immunomodulation potential and ability to differentiate into multi-lineages, which makes them a tool of interest in cell therapy. Toxicological, immunological and contamination risks are concerns in the culture of cells for cell therapy, and may be associated with the use of animal supplements, such as fetal bovine serum (FBS). Human cell cultures have been successful in supplementing human serum in substitution for FBS, which may address problems eventually attributed to the use of non-species-specific supplements. In parallel, studies with extracellular vesicles (EVs) have demonstrated great potential of these particles in cell culture, being able, for example, to induce the differentiation of stem cells. This project aims to evaluate the potential of extracellular vesicles derived from pooled adult human serum (HS) regarding the ability to support the culture of human MSCs derived from adipose tissue (n = 3). FBS, HS, and extracellular vesicles-depleted adult human serum (DS) were also evaluated. The groups were evaluated for proliferation, viability, senescence and cell differentiation. Extracellular vesicles from human serum were not able to maintain the proliferation and viability potential of MSCs compared to the other groups. Among FBS, HS and DS groups, cells cultured with human serum and depleted human serum had higher proliferative potential in the first passages, and those cultured with FBS at later passages, but cell viability at all passages was higher in cells cultured with FBS. In relation to cell senescence, there was no difference between cells cultured with FBS and HS, but culture with DS resulted in a lower percentage of senescent cells. All sera maintained chondrogenic, osteogenic and adipogenic potential of MSCs. In addition, differences in the response of different MSCs isolates were observed in relation to serum types, indicating that the origin of the cells influenced their behavior and suggesting the need for studies with a higher number of isolates. Finally, while other studies have shown that complete human serum is an appropriate substitute for FBS, this study showed that DS is also a possible substitute for fetal bovine serum, with potential use in culturing MSCs to isolate EVs produced by these cells. In particular, depleted human serum stood out for having fewer senescent cells than the other groups.
• Link de acesso: https://bdtd.ucb.br:8443/jspui/handle/tede/2556

Mesenchymal stem cells (MSCs) have immunomodulation potential and ability to differentiate into multi-lineages, which makes them a tool of interest in cell therapy. Toxicological, immunological and contamination risks are concerns in the culture of cells for cell therapy, and may be associated with the use of animal supplements, such as fetal bovine serum (FBS). Human cell cultures have been successful in supplementing human serum in substitution for FBS, which may address problems eventually attributed to the use of non-species-specific supplements. In parallel, studies with extracellular vesicles (EVs) have demonstrated great potential of these particles in cell culture, being able, for example, to induce the differentiation of stem cells. This project aims to evaluate the potential of extracellular vesicles derived from pooled adult human serum (HS) regarding the ability to support the culture of human MSCs derived from adipose tissue (n = 3). FBS, HS, and extracellular vesicles-depleted adult human serum (DS) were also evaluated. The groups were evaluated for proliferation, viability, senescence and cell differentiation. Extracellular vesicles from human serum were not able to maintain the proliferation and viability potential of MSCs compared to the other groups. Among FBS, HS and DS groups, cells cultured with human serum and depleted human serum had higher proliferative potential in the first passages, and those cultured with FBS at later passages, but cell viability at all passages was higher in cells cultured with FBS. In relation to cell senescence, there was no difference between cells cultured with FBS and HS, but culture with DS resulted in a lower percentage of senescent cells. All sera maintained chondrogenic, osteogenic and adipogenic potential of MSCs. In addition, differences in the response of different MSCs isolates were observed in relation to serum types, indicating that the origin of the cells influenced their behavior and suggesting the need for studies with a higher number of isolates. Finally, while other studies have shown that complete human serum is an appropriate substitute for FBS, this study showed that DS is also a possible substitute for fetal bovine serum, with potential use in culturing MSCs to isolate EVs produced by these cells. In particular, depleted human serum stood out for having fewer senescent cells than the other groups.