Descongelamento de organóides intestinais caninos : técnicas e desafios
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2023
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Os organóides são modelos de cultura celular tridimensionais complexos, formados a partir de células tronco pluripotentes ou de células mãe, com capacidade de auto-organização e que replicam a fisiologia in vivo de uma maneira muito mais precisa do que os sistemas de cultura celular bidimensionais. Existe um grande número de aplicações para os organóides incluindo estudos sobre organogénese, desenvolvimento de novos fármacos, autorrenovação celular, medicina regenerativa, patogénese do cancro e de outras patologias crónicas como é o caso da Inflamatory Bowell Desease nos cães. Através do método de criopreservação é possível manter culturas de organóides viáveis durante meses ou anos com a possibilidade de se descongelarem quando se pretender. Neste trabalho desenvolveram-se dois protocolos de descongelação de organóides caninos e analisou-se o desenvolvimento dos mesmos nas primeiras 72 horas pós descongelamento. Avaliou-se ainda a viabilidade de o uso de um meio humano nos enteróides caninos. Concluiu-se que é possível criopreservar organóides durante anos, conservando a sua viabilidade, apesar de ainda existirem muitas limitações associadas ao processo de descongelamento dos mesmos. Palavras-chave: Criopreservação, Culturas, Descongelação, Organóides, Pluripotentes
Organoids are complex three-dimensional cell culture models, formed from pluripotent stem cells or stem cells, with the capacity for self-organization and replication of in vivo physiology in a much more precise way than two-dimensional cell culture systems. There is a wide range of applications for organoids, including studies on organogenesis, drug development, cell self-renewal, regenerative medicine, cancer pathogenesis, and other chronic pathologies, such as Inflammatory Bowel Disease in dogs. Through the method of cryopreservation, it is possible to maintain viable organoid cultures for months or years with the possibility of thawing them when desired. In this study, two canine organoid thawing protocols were developed, and their development was analyzed in the first 72 hours post-thawing. The feasibility of using a human medium for canine enteroids was also assessed. It was concluded that it is possible to cryopreserve organoids for years while preserving their viability, although there are still many limitations associated with the thawing process. Keywords: Cryopreservation, Cultures, Organoids, Pluripotent, Thawing
Organoids are complex three-dimensional cell culture models, formed from pluripotent stem cells or stem cells, with the capacity for self-organization and replication of in vivo physiology in a much more precise way than two-dimensional cell culture systems. There is a wide range of applications for organoids, including studies on organogenesis, drug development, cell self-renewal, regenerative medicine, cancer pathogenesis, and other chronic pathologies, such as Inflammatory Bowel Disease in dogs. Through the method of cryopreservation, it is possible to maintain viable organoid cultures for months or years with the possibility of thawing them when desired. In this study, two canine organoid thawing protocols were developed, and their development was analyzed in the first 72 hours post-thawing. The feasibility of using a human medium for canine enteroids was also assessed. It was concluded that it is possible to cryopreserve organoids for years while preserving their viability, although there are still many limitations associated with the thawing process. Keywords: Cryopreservation, Cultures, Organoids, Pluripotent, Thawing
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VETERINARY MEDICINE, CRYOPRESERVATION, DOGS, ORGANOIDS, MESTRADO INTEGRADO EM MEDICINA VETERINÁRIA, VETERINÁRIA, MEDICINA VETERINÁRIA, CRIOPRESERVAÇÃO, CÃES, ORGANÓIDES, Mestrado Integrado em Medicina Veterinária