RESUMEN
Docosahexaenoic acid (DHA) is an n-3 polyunsaturated fatty acid (PUFA) that improves fertility by increasing membrane fluidity. Moreover, embryos produced by donor females supplied with n-3 PUFA did not show any difference in terms of the lipid profile after 7 days of culture. The present study aimed to investigate the effects of DHA (20 and 100 µM) coupled with carnosine (5 mg/mL), an antioxidant, during oocyte maturation and embryo development on the developmental and cryosurvival rates and the number of pluripotent cells. Free fatty acid receptor-4 (FFAR4), which is able to bind DHA, was visualised by immunostaining. The addition of DHA in the in vitro development (IVD) medium decreased the percentage of pluripotent SOX2 positive cells compared with the control (8.4% vs. 10.9%) without affecting the number of cells (196.7 vs. 191.6 cells) or the developmental (20.9% vs. 23.9% blastocysts rate on D7) and cryosurvival rates (86.3% vs 86.2%). Such a decrease in pluripotent cells, relevant to the differentiation of the first lineage within the inner cell mass, represents an improvement in the embryo quality. On the contrary, embryos without any pluripotent SOX2-positive cells would not be able to achieve gestation. Future studies should follow up these results by carrying out embryo transfers to assess the beneficial effects of DHA supplementation.
Asunto(s)
Ácidos Docosahexaenoicos , Técnicas de Maduración In Vitro de los Oocitos , Animales , Blastocisto , Bovinos , Criopreservación/veterinaria , Ácidos Docosahexaenoicos/farmacología , Técnicas de Cultivo de Embriones/veterinaria , Desarrollo Embrionario , Femenino , Fertilización In Vitro/veterinaria , Técnicas de Maduración In Vitro de los Oocitos/métodos , Técnicas de Maduración In Vitro de los Oocitos/veterinaria , OocitosRESUMEN
Rabbit induced pluripotent stem cells (rbiPSCs) possess the characteristic features of primed pluripotency as defined in rodents and primates. In the present study, we reprogrammed rbiPSCs using human Krüppel-like factors (KLFs) 2 and 4 and cultured them in a medium supplemented with fetal calf serum and leukemia inhibitory factor. These cells (designated rbEKA) were propagated by enzymatic dissociation for at least 30 passages, during which they maintained a normal karyotype. This new culturing protocol resulted in transcriptional and epigenetic reconfiguration, as substantiated by the expression of transcription factors and the presence of histone modifications associated with naïve pluripotency. Furthermore, microarray analysis of rbiPSCs, rbEKA cells, rabbit ICM cells, and rabbit epiblast showed that the global gene expression profile of the reprogrammed rbiPSCs was more similar to that of rabbit ICM and epiblast cells. Injection of rbEKA cells into 8-cell stage rabbit embryos resulted in extensive colonization of ICM in 9% early-blastocysts (E3.5), epiblast in 10% mid-blastocysts (E4.5), and embryonic disk in 1.4% pre-gastrulae (E6). Thus, these results indicate that KLF2 and KLF4 triggered the conversion of rbiPSCs into epiblast-like, embryo colonization-competent PSCs. Our results highlight some of the requirements to achieve bona fide chimeric competency.
Asunto(s)
Reprogramación Celular , Estratos Germinativos/citología , Células Madre Pluripotentes Inducidas/citología , Factores de Transcripción de Tipo Kruppel/metabolismo , Animales , Blastocisto/citología , Blastocisto/metabolismo , Proliferación Celular , Supervivencia Celular , Quimera/metabolismo , Epigénesis Genética , Perfilación de la Expresión Génica , Humanos , Factor 4 Similar a Kruppel , Ratones , MicroARNs/genética , MicroARNs/metabolismo , Conejos , Transducción de SeñalAsunto(s)
Informes de Casos , Humanos , Masculino , Femenino , Lactante , Preescolar , Niño , Adolescente , Arsenicum Album , Síntomas PsíquicosRESUMEN
Crier, pour un bebe, n'est-ce pas aussi normal, aussi indispensable que respirer? Sans le premier cri, pas de deplissement des alveoles pulmonaires, pas de passage de la vie aquatique a la vie aerienne... (AU)