In vitro production of porcine embryos with use of chemically semi-defined culture media system

The objective of this study was to determine the effect of a semi-defined culture media system developed in our laboratory, named Pigs Media System (PMS) on the in vitro production of porcine embryos. In a first assay, the cummulus-oocytes complexes (COCs) were matured, fertilized and cultured for embryo development in PMS supplemented with bovine serum albumin (BSA), and in North Carolina State University-23 (NCSU-23) supplemented with fluid follicular, until blastocysts evaluation. In the assay 2, maturation and culture were performed in PMS using BSA or polyvinyl alcohol (PVA) in a 2 × 2 factorial arrangement (PMS-BSA/BSA, PMS-BSA/PVA, PMS-PVA/PVA, PMS-PVA/BSA). The PMS had a positive effect on the total cell number (58.04) and the decrease of the total lipids (49.4%) regarding the NCSU-23 medium (37.98 and 59.2% respectively; p<0.05). The percentage of monospermic fertilization was significantly lower (42.3%; p<0.05) when oocytes were matured with PMS-BSA than in PMS-PVA (52.6%). The supplementation of BSA in the PMS for embryo culture, increased the blastocyst development, the cell number of blastocysts and decreased the content of total lipids (36.8%, 46.9 and 49.6% respectively; p<0.05), in comparison with the supplementation of PVA in the PMS for embryo culture. These results suggest that the semi-defined culture media system developed by the National Genetic Resources Center (CNRG), have proved favorable effects on the total cell number and the decrease of total lipids of porcine blastocysts in vitro produced. The objective of this study was to determine the effect of a semi-defined culture media system developed in our laboratory, named Pigs Media System (PMS) on the in vitro production of porcine embryos. In a first assay, the cummulus-oocytes complexes (COCs) were matured, fertilized and cultured for embryo development in PMS supplemented with bovine serum albumin (BSA), and in North Carolina State University-23 (NCSU-23) supplemented with fluid follicular, until blastocysts evaluation. In the assay 2, maturation and culture were performed in PMS using BSA or polyvinyl alcohol (PVA) in a 2 × 2 factorial arrangement (PMS-BSA/BSA, PMS-BSA/PVA, PMS-PVA/PVA, PMS-PVA/BSA). The PMS had a positive effect on the total cell number (58.04) and the decrease of the total lipids (49.4%) regarding the NCSU-23 medium (37.98 and 59.2% respectively; p<0.05). The percentage of monospermic fertilization was significantly lower (42.3%; p<0.05) when oocytes were matured with PMS-BSA than in PMS-PVA (52.6%). The supplementation of BSA in the PMS for embryo culture, increased the blastocyst development, the cell number of blastocysts and decreased the content of total lipids (36.8%, 46.9 and 49.6% respectively; p<0.05), in comparison with the supplementation of PVA in the PMS for embryo culture. These results suggest that the semi-defined culture media system developed by the National Genetic Resources Center (CNRG), have proved favorable effects on the total cell number and the decrease of total lipids of porcine blastocysts in vitro produced.

Biotecnologías reproductivas en el ganado bovino: cinco décadas de investigación en México

The main bovine reproductive biotechnologies are recapitulated herein in five sections, and their historical development and current status are analyzed, including the results generated in Mexico. In the 1970s, estrus synchronization and ovulation induction began; thus, the reproductive cycle started to be controlled with the resources available at that time, based on the knowledge of bovine reproductive physiology. Over the years, hormone therapy evolved as new compounds were discovered, refining methods to standardize the effect and generating new methods for the release of hormones. The most widely used biotechnology in the world, artificial insemination, owes its expansion to advances in semen processing, among which the development of diluents, cryopreservation, semen sexing, and computer-assisted sperm analysis stand out. The embryonic era began with the development of multi-ovulation and methods for collecting, evaluating, transferring, and cryopreserving embryos. The second embryonic era came with the fully in vitro production of embryos from immature eggs and frozen sperm, known as in vitro embryo production. Great research and material resources have been invested in this procedure, rendering it a pillar of genetic improvement and productivity, in combination with two other tools: sexed semen and genomic evaluations. A golden age of in vitro embryo production is on the horizon, with the possibility to produce accurate modifications in the embryo genome, thanks to gene editing technology.A lo largo de cinco secciones, se recapitulan las principales biotecnologías reproductivas en el bovino, se analiza su desarrollo histórico, estado actual, y se incluyen resultados generados en México. En la década de 1970, se inició la sincronización estral e inducción de la ovulación donde, basados en el conocimiento de la fisiología reproductiva bovina, se empezó a controlar el ciclo reproductivo con recursos disponibles en aquel entonces. Con los años, la terapia hormonal evolucionó conforme se descubrieron nuevos compuestos, refinando métodos para estandarizar el efecto y generar nuevos métodos de liberación de las hormonas. La biotecnología más usada en el mundo, la inseminación artificial, debe su expansión a los avances en el procesamiento del semen, donde destaca el desarrollo de diluyentes, la criopreservación, el sexado del semen y el análisis espermático asistido por computadora. La era embrionaria inició con el desarrollo de la multiovulación y los métodos para colectar, evaluar, transferir y criopreservar los embriones. La segunda era embrionaria llegó con la producción de embriones completamente in vitro, partiendo de óvulos inmaduros y semen congelado, en lo que se denominó la producción in vitro de embriones. En ésta, se han invertido grandes recursos de investigación, y materiales, para hacerla un pilar del mejoramiento genético y la productividad, en combinación con otras dos herramientas, el semen sexado y las evaluaciones genómicas. Se vislumbra una época de oro de la producción in vitro de embriones con la posibilidad de modificar el genoma de embriones con precisión, gracias a la tecnología de edición de genes

Caracterización de la respuesta ovárica a la superovulación en bovino Criollo Coreño utilizando dosis reducidas de FSH

To evaluate the ovarian response to superovulation using reduced doses of FSH in Criollo Coreño cattle two experiments were conducted at the Experimental Station “El Verdineño” (INIFAP) located in Nayarit, México. Twelve (12) cows of 12.4 ± 2.9 yr old (Exp 1) and six heifers of 3.0 ± 0.3 yr old (Exp 2) were used. Three treatments were assigned to each female consisting of 280 mg (T1), 200 mg (T2) and 140 mg (T3) of FSH, so that all females received all treatments. The response variables were transferable embryos (ET), corpuscles retrieved (CR) = (embryos+non fertilized ova), degenerated embryos (ENT), number of corpora lutea (CL), non-fertilized ova (ONF), ovarian volume (VO), serum progesterone (P4), fertilization rate (%F) = ((ET+ENT)*100)/CR) and percent recovery (%R)=CL*100/CR. A cross-over balanced experimental design was used. Analyses were carried out with the GLM procedure of SAS. The statistical model included the fixed effects of treatment (T1, T2 and T3), animal (twelve or six) and period (three periods). In Exp 1 differences among treatments (P0.05) among treatments for ET, ENT, CL, VO, P4, %F or %R. In Exp 2 no differences were detected (P>0.05) among treatments for any of the variables. It is feasible to use reduced doses of FSH for the superovulation of Criollo Coreño heifers without affecting the response to superovulation or to embryo production. Para evaluar la respuesta ovárica a la superovulación con dosis reducidas de la hormona folículo estimulante (FSH) en bovinos Criollo Coreño, se realizaron dos experimentos en el Sitio Experimental El Verdineño del INIFAP ubicado en Nayarit, México. Se utilizaron 12 vacas de 12.4 ± 2.9 años (Exp 1) y seis vaquillas de 3.0 ± 0.3 años (Exp 2). Se aplicaron tres tratamientos a cada hembra: 280 mg (T1), 200 mg (T2) y 140 mg (T3) de FSH. Las variables evaluadas fueron número de embriones transferibles (ET), número de corpúsculos recuperados (CR)=(embriones + óvulos no fertilizados), número de embriones no transferibles (ENT), número de cuerpos lúteos (CL), número de óvulos no fertilizados (ONF), volumen ovárico (VO), concentración sérica de progesterona (P4), porcentaje de fertilización (%F)=((ET+ENT)*100)/CR) y porcentaje de recuperación (%R)=CL*100/CR. Se utilizó un diseño experimental crossover simple balanceado. La información se analizó con el procedimiento GLM de SAS. El modelo estadístico incluyó los efectos de tratamiento (T1, T2 y T3), vaquilla/vaca (6 y 12) y período (3 períodos). En el Exp 1 se detectaron diferencias (P0.05) entre tratamientos para ET, ENT, CL, VO, P4, %F o %R. En el Exp 2 no se detectaron diferencias (P>0.05) entre tratamientos para ninguna de las variables. Es factible utilizar dosis reducidas de FSH para la superovulación en vaquillas Criollo Coreño sin afectar la respuesta a la superovulación o la producción de embriones.

Vitrification of White-tailed Deer (Odocoileus virginianus) oocytes with sucrose or trehalose for in vitro maturation and fertilization.

Objective: Evaluate the White-tailed Deer (WTD) in vitro embryo production (IVP) and oocytes vitrified with Trehalose (TH) or Sucrose (SC). Design/methodology/approach: Total vitrified oocytes were placed into two different groups: TH (n=60) and SC (n=61). Samples were selected and analyzed for viability evaluation TH (n=5) and SC (n=5), nuclear status (NS) TH (n=4) and SC (n=5), Germinal Vesicle (GV), Metaphase I, or not evaluable (NE) after warming. In vitro maturation (IVM) was conducted for 36 h in supplemented TCM-199 medium. Immediately afterwards, oocyte NS was evaluated (n=88) [(GV, MI=immature), (MII=mature)]. In vitro fertilization (IVF) was performed in supplemented TALP medium for 24 h using frozen WTD semen (3x106 sperm/mL), NS was classified [Fertilized (F), Not fertilized (NF), or NE]. Results: After warming, viability for the TH group (n=5) was 60% versus 40% for SC group (n=5), however, oocytes in both groups were immature (GV and MI stage). For IVM, NS evaluations of the TH group (n=38) revealed no maturation versus 2% in the SC group (n=50) (MII stage=matured). IVF evaluations for the TH group (n=10) revealed no fertilization compared to 20% in the SC group (n=5). A statistical difference (p&gt;0.05) was not found between the TH and SC groups. Limitations on study/implications: White-tailed Deer in vitro embryo production is not well documented. Findings/conclusions: Future research with a larger number of WTD oocytes is needed for further evaluation of oocyte vitrification IVP techniques as a model for endangered cervids

Escuela secundaria posible

REPENSAR LA EDUCACIÓN SECUNDARIA Colección de infografías II Parte Seguimiento de medios de comunicación 2017-2019Fil: Ferreyra, Horacio Ademar. Universidad Católica de Córdoba. Facultad de Educación; ArgentinaFil: Di Francesco, Adriana Carlota. Universidad Católica de Córdoba. Facultad de Educación; ArgentinaFil: Equipo de investigación en educación de adolescentes y jóvenes. Unidad Asociada CONICET. Universidad Católica de Córdoba. Facultad de Educación; Argentina