RESUMO
This study aimed to demonstrate the applicability of the ovarian slicing technique in the recovery of post mortem oocytes in a llama (lama glama) victim of acute bronchopneumonia and the effectiveness of the brilliant cresyl blue (ACB) technique to select quality oocytes for a subsequent in vitro culture. The ovary-salpinge-hysterectomy (OSH) was performed 12 hours after the animal's death. The ovaries were transported refrigerated immediately to the laboratory in Leibovitz medium (L-15). The oocytes were identified, quantified under a stereomicroscopic and classified according to two methods: conventional morphological classification and ACB assay. 26 oocytes classified by conventional morphology were obtained: 10 oocytes grade 1, 12 oocytes grade 2 and 4 oocytes grade3. The use of the ACB dye demonstrated that 88% of the evaluated oocytes maintained their quality and were competent for the conservation of genetic or future material application in reproductive biotechnologies in this species. Further studies are suggested aiming to use selected oocytes with ACB to enhance the success of in vitro culture in this species.(AU)
Assuntos
Animais , Feminino , Camelídeos Americanos/genética , Técnicas de Maturação in Vitro de Oócitos/métodos , Técnicas de Maturação in Vitro de Oócitos/veterinária , Oócitos/efeitos dos fármacos , Reprodução/efeitos dos fármacosRESUMO
This study aimed to demonstrate the applicability of the ovarian slicing technique in the recovery of post mortem oocytes in a llama (lama glama) victim of acute bronchopneumonia and the effectiveness of the brilliant cresyl blue (ACB) technique to select quality oocytes for a subsequent in vitro culture. The ovary-salpinge-hysterectomy (OSH) was performed 12 hours after the animal's death. The ovaries were transported refrigerated immediately to the laboratory in Leibovitz medium (L-15). The oocytes were identified, quantified under a stereomicroscopic and classified according to two methods: conventional morphological classification and ACB assay. 26 oocytes classified by conventional morphology were obtained: 10 oocytes grade 1, 12 oocytes grade 2 and 4 oocytes grade3. The use of the ACB dye demonstrated that 88% of the evaluated oocytes maintained their quality and were competent for the conservation of genetic or future material application in reproductive biotechnologies in this species. Further studies are suggested aiming to use selected oocytes with ACB to enhance the success of in vitro culture in this species.
Assuntos
Feminino , Animais , Camelídeos Americanos/genética , Oócitos/efeitos dos fármacos , Reprodução/efeitos dos fármacos , Técnicas de Maturação in Vitro de Oócitos/métodos , Técnicas de Maturação in Vitro de Oócitos/veterináriaRESUMO
O objetivo do estudo foi avaliar a viabilidade e taxa de proliferação de células-tronco mesenquimais (CTMs) derivadas do líquido amniótico (LA) após cultivo in vitro, e o efeito de dois agentes crioprotetores. Foram utilizadas 9 cabras prenhes. As amostras foram colhidas de fetos caprinos por laparotomia, e delas obtidos as (CTMs) e submetidas ao cultivo in vitro. Posteriormente, uma fração das células foi criopreservada em meio DMSO (dimetilsulfóxido) ou glicerol e vitrificadas para posterior avaliação da viabilidade. O meio DMSO promoveu melhores taxas de sobrevida celular preservando as características de pluripotencialidade e de replicação in vitro.(AU)
The objective of the study was to evaluate the viability and proliferation rate of mesenchymal stem cells (MTCs) derived from amniotic fluid (LA) after in vitro culture, and the effect of two cryoprotective agents. 9 pregnant goats were used. The samples were collected from goat fetuses by laparotomy, and from them (CTMs) were obtained and cultured in vitro. Subsequently, a fraction of the cells were cryopreserved in DMSO (dimethylsulfoxide) or glycerol medium and vitrified for further evaluation of viability. The DMSO medium promoted better cell survival rates while preserving the characteristics of pluripotency and replication in vitro.(AU)
Assuntos
Animais , Feminino , Ruminantes , Células-Tronco/fisiologia , Líquido Amniótico , Crioprotetores/análise , Dimetil Sulfóxido/análise , Glicerol/análise , Técnicas In Vitro/veterinária , Técnicas In Vitro/métodosRESUMO
O objetivo do estudo foi avaliar a viabilidade e taxa de proliferação de células-tronco mesenquimais (CTMs) derivadas do líquido amniótico (LA) após cultivo in vitro, e o efeito de dois agentes crioprotetores. Foram utilizadas 9 cabras prenhes. As amostras foram colhidas de fetos caprinos por laparotomia, e delas obtidos as (CTMs) e submetidas ao cultivo in vitro. Posteriormente, uma fração das células foi criopreservada em meio DMSO (dimetilsulfóxido) ou glicerol e vitrificadas para posterior avaliação da viabilidade. O meio DMSO promoveu melhores taxas de sobrevida celular preservando as características de pluripotencialidade e de replicação in vitro.
The objective of the study was to evaluate the viability and proliferation rate of mesenchymal stem cells (MTCs) derived from amniotic fluid (LA) after in vitro culture, and the effect of two cryoprotective agents. 9 pregnant goats were used. The samples were collected from goat fetuses by laparotomy, and from them (CTMs) were obtained and cultured in vitro. Subsequently, a fraction of the cells were cryopreserved in DMSO (dimethylsulfoxide) or glycerol medium and vitrified for further evaluation of viability. The DMSO medium promoted better cell survival rates while preserving the characteristics of pluripotency and replication in vitro.