Impact of high-fat diet consumption during prolonged period of pregnancy on placenta structures and umbilical vascular growth in goats

This study aimed to verify the impact of high-fat diet consumption for a prolonged period on oxidative stress, fetal growth, umbilical vascular system, and placental structures in pregnant goats. Twenty-two pregnant goats were grouped into the control diet (n= 11) and fat diet (n = 11). Flaxseed meal was added to the fat diet, replacing the corn grain of concentrate, from gestational day 100 to delivery date. Diets were isonitrogenous and isoenergetic, differing in fat content (2.8% vs. 6.3% dry matter). The fat group showed higher feed intake and total plasma lipid levels than the control group (P < 0.001). No difference was found in placentome, and umbilical vascular development. Fat diet-fed goats exhibited a lower systolic peak in the umbilical artery. At delivery, placental traits were similar with the exception of the cotyledon width (P = 0.0075), which was smaller in the fat group and cotyledon surface (P = 0.0047) for multiple pregnancy of fat diet. Cotyledonary epithelium showed more intense staining of lipid droplets and a greater area for lipofuscin staining in the fat group compared to control group (P < 0.001). The mean live weight of the kids was lower in the fat group in the first week after delivery than in control group. Thus, in goats, the continuous administration of a high-fat diet during pregnancy does not appear to modify the fetal-maternal vascular structures but has an impact on a part of the placental structure; therefore, its use must be carefully evaluated.(AU)

Glycerin supplementation strategies for three or seven days affects oxidative stress, follicle dynamics and ovulatory response in Morada Nova sheep

The Ziwuling black goat is an indigenously in China, their offspring are frequently affected by congenital cryptorchidism. The extracellular matrix (ECM) contains cytokines and growth factors that regulate the development of the testis, and component changes often result in pathological changes. Cryptorchidism is closely related to structural changes in ECM. In this study, the histochemical staining, immunohistochemical, immunofluorescence and Western blot combined with semi-quantitative analysis was used to describe the distribution of the important ECM components Collagen type IV (Col IV), laminin (LN)and heparan sulfate proteoglycans (HSPG) in the normal and cryptorchid testes of Ziwuling black goats. Results showed that: The histochemical staining showed that the dysplasia of seminiferous tubules and decreased number of Sertoli cells in cryptorchidism, as well as sparse collagen fiber. Meanwhile, the distribution of reticular fibers is relatively rich. Furthermore, the PAS and AB staining in the interstitial vessels and lamina propria of seminiferous tubules is weak. The immunohistochemical and immunofluorescence revealed that Col IV, LN was strongly expressed in Leydig, Sertoli cells of normal testes and moderately positive in the spermatogonia and spermatids, but HSPG was not expressed in the spermatogonia. However, cryptorchidism, the expression of Col IV, LN and HPSG in Leydig, Sertoli cells significantly decreased, as well as the expression of Col IV and LN in capillary endothelial cells, but HSPG was moderately expressed in spermatogonia. Based on these data, the underdevelopment of spermatogenic epithelium, decreased synthesis function of collagen fibers and Leydig cells develop usually in the cryptorchidism were shown to be closely related to the abnormal metabolism of Col IV and LN. The positive expressed of HSPG in the spermatogonia of cryptorchid testes is related to the compensatory development of spermatogonia.(AU)

Fluido amniótico, alternativa eficiente para obtenção de células-tronco mesenquimais multipotentes / Amniotic fluid, efficient alternative to obtain multipotent mesenchymal stem cells

Na última década houve um crescente interesse na investigação quanto à presença de células-tronco no fluido amniótico, devido a facilidade de obtenção, isolamento e cultivo in vitro. Além disso, a utilização do fluido como fonte de células-tronco possibilita a obtenção de células com alto grau de indiferenciação e elevada taxa de proliferação in vitro e grande capacidade de diferenciação. Todas estas vantagens tornam o líquido amniótico uma fonte atrativa de células-tronco para utilização em biotecnologias reprodutivas, como a clonagem e transgênese, bem como ensaios clínicos e terapias celulares em animais para posterior utilização em seres humanos. Este artigo apresenta um panorama da pesquisa científica com células-tronco derivadas do fluido amniótico no mundo, a partir de levantamento bibliográfico de artigos científicos de pesquisadores brasileiros e estrangeiros.

In the last decade there has been a growing interest in investigation of presence of stem cells in amniotic fluid due to the ease of obtaining, isolating and in vitro culture. In addition, the use of amniotic fluid as a source of stem cells makes it possible to obtain cells with a high degree of indifferentiation and a high rate of in vitro proliferation and a great capacity for differentiation. All of these advantages make amniotic fluid an attractive source of stem cells for use in reproductive biotechnologies, such as cloning and transgenesis, as well as clinical trials in animals for further use in humans. This article presents a panorama of the scientific research with stem cells derived from amniotic fluid in the world, from a bibliographical survey of scientific articles of Brazilian and foreign researchers.

Fluido amniótico, alternativa eficiente para obtenção de células-tronco mesenquimais multipotentes / Amniotic fluid, efficient alternative to obtain multipotent mesenchymal stem cells

Na última década houve um crescente interesse na investigação quanto à presença de células-tronco no fluido amniótico, devido a facilidade de obtenção, isolamento e cultivo in vitro. Além disso, a utilização do fluido como fonte de células-tronco possibilita a obtenção de células com alto grau de indiferenciação e elevada taxa de proliferação in vitro e grande capacidade de diferenciação. Todas estas vantagens tornam o líquido amniótico uma fonte atrativa de células-tronco para utilização em biotecnologias reprodutivas, como a clonagem e transgênese, bem como ensaios clínicos e terapias celulares em animais para posterior utilização em seres humanos. Este artigo apresenta um panorama da pesquisa científica com células-tronco derivadas do fluido amniótico no mundo, a partir de levantamento bibliográfico de artigos científicos de pesquisadores brasileiros e estrangeiros.(AU)

In the last decade there has been a growing interest in investigation of presence of stem cells in amniotic fluid due to the ease of obtaining, isolating and in vitro culture. In addition, the use of amniotic fluid as a source of stem cells makes it possible to obtain cells with a high degree of indifferentiation and a high rate of in vitro proliferation and a great capacity for differentiation. All of these advantages make amniotic fluid an attractive source of stem cells for use in reproductive biotechnologies, such as cloning and transgenesis, as well as clinical trials in animals for further use in humans. This article presents a panorama of the scientific research with stem cells derived from amniotic fluid in the world, from a bibliographical survey of scientific articles of Brazilian and foreign researchers. (AU)

Efeito do DMSO e glicerol na criopreservação de células tronco mesenquimais derivadas de líquido amniótico de fetos caprinos / Effect of DMSO and glycerol in criopreservation of mesenquimial trunk cells derived from amniotic fluid of caprine fetus

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.

Efeito do escore da condição corporal materna no desenvolvimento in vitro das células tronco mesenquimais do cordão umbilical de fetos caprinos / Effect of the score of maternal body condition in the in vitro development of the mesenquimal trunk cells in the umbilical cord of caprine fetes

O objetivo do trabalho foi avaliar o efeito do escore da condição corporal (ECC) materno sobre a capacidade de isolamento, expansão e caracterização de células-tronco mesenquimais (CTMs) derivadas do cordão umbilical de fetos caprinos. Para tanto, dezenove cabras mestiças adultas, pluríparas foram agrupadas de acordo com o ECC, atribuindo-se um escore de 1-5, (GB) grupo baixo com menor ECC, (2,3±0,1, GB, n = 9) e (GA) grupo alto com maior ECC (2,9±0,1, GA, n = 10). Durante o parto, fragmentos do cordão umbilical foram coletados e cultivados in vitro e avaliados a taxa de proliferação celular. Nenhum efeito significativo do ECC foi encontrado para os parâmetros considerados.

The objective of this study was to evaluate the effect of the maternal body condition score (ECC) on the ability to isolate, expand and characterize mesenchymal stem cells (MTCs) derived from the umbilical cord of goat fetuses. Nineteen crossbred adult crossbred goats were grouped according to ECC, giving a score of 1-5 (GB) low group with lower ECC, (2.3±0.1, GB, n = 9 ) and (GA) high group with higher ECC (2.9±0.1, GA, n = 10). During delivery, umbilical cord fragments were collected and cultured in vitro and the rate of cell proliferation was evaluated. No significant effect of ECC was found for the parameters considered.

Efeito do DMSO e glicerol na criopreservação de células tronco mesenquimais derivadas de líquido amniótico de fetos caprinos / Effect of DMSO and glycerol in criopreservation of mesenquimial trunk cells derived from amniotic fluid of caprine fetus

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)

Efeito do escore da condição corporal materna no desenvolvimento in vitro das células tronco mesenquimais do cordão umbilical de fetos caprinos / Effect of the score of maternal body condition in the in vitro development of the mesenquimal trunk cells in the umbilical cord of caprine fetes

O objetivo do trabalho foi avaliar o efeito do escore da condição corporal (ECC) materno sobre a capacidade de isolamento, expansão e caracterização de células-tronco mesenquimais (CTMs) derivadas do cordão umbilical de fetos caprinos. Para tanto, dezenove cabras mestiças adultas, pluríparas foram agrupadas de acordo com o ECC, atribuindo-se um escore de 1-5, (GB) grupo baixo com menor ECC, (2,3±0,1, GB, n = 9) e (GA) grupo alto com maior ECC (2,9±0,1, GA, n = 10). Durante o parto, fragmentos do cordão umbilical foram coletados e cultivados in vitro e avaliados a taxa de proliferação celular. Nenhum efeito significativo do ECC foi encontrado para os parâmetros considerados.(AU)

The objective of this study was to evaluate the effect of the maternal body condition score (ECC) on the ability to isolate, expand and characterize mesenchymal stem cells (MTCs) derived from the umbilical cord of goat fetuses. Nineteen crossbred adult crossbred goats were grouped according to ECC, giving a score of 1-5 (GB) low group with lower ECC, (2.3±0.1, GB, n = 9 ) and (GA) high group with higher ECC (2.9±0.1, GA, n = 10). During delivery, umbilical cord fragments were collected and cultured in vitro and the rate of cell proliferation was evaluated. No significant effect of ECC was found for the parameters considered.(AU)

Comparative expression profiles of genes related to cellular stress and development in oocyte of goats fed with distinct concentrations of lipids

Background: Lipotoxicity is characterized by an excess of saturated fatty acids in the blood stream, in which other nonadipose cells begin to store them, thereby altering the expression of genes related to endoplasmic reticulum stress, whoseeffects have been associated with decreased oocyte quality in several species, decreased mitochondrial activity, and increasedapoptosis. The present study was conducted to investigate the lipotoxicity effect of diets with increasing fat levels on geneexpression in goats oocyte and granulosa cells.Materials, Methods & Results: Thirty does were divided into three groups of 10 animals each, which received forage andconcentrate to provide respectively 2.7% of lipids (LL group), 3.9% lipids (LI group) and 5.1% lipids (LH group) for 28days. Three days before oocyte harvest, follicular wave was synchronized by 1 mL PGF2α intramuscularly, followed bythe insertion of an intravaginal progesterone release device. Viable oocytes and granulosa cells were subjected to qRT-PCRto determine the expression of PLIN2, ATF4, CHOP10, BAX, BCL2, HSP70 genes, were checked, evaluated using thedissociation curve analysis, which obtained the following values: 77.8, 80.2, 83.6, 78.0, 81.0 and 82.0ºC, respectively. Thesteps of qPCR thermal cycle was: denaturation and polymerase activation, annealing and final extension.Throughout theexperimental period, blood samples were taken for cholesterol, triglycerides and total lipids measurements. Total plasmalipids determination was calculated with the following equation: 2 x (cholesterol + triglycerides) × 1.1 with sensitivity ofthe assay for cholesterol was 1.472 mg/dL and for triglycerides, 2.845 mg/dL. In LH group, it was recorded a more pronounced increase in total lipids (+ 60.1%) (P < 0.05), cholesterol (+ 18.8%) and triglycerides (+ 8.5%). These increaseswere three times higher than that in LL and LI groups. All genes were expressed in oocytes...(AU)

Comparative expression profiles of genes related to cellular stress and development in oocyte of goats fed with distinct concentrations of lipids

Background: Lipotoxicity is characterized by an excess of saturated fatty acids in the blood stream, in which other nonadipose cells begin to store them, thereby altering the expression of genes related to endoplasmic reticulum stress, whoseeffects have been associated with decreased oocyte quality in several species, decreased mitochondrial activity, and increasedapoptosis. The present study was conducted to investigate the lipotoxicity effect of diets with increasing fat levels on geneexpression in goat’s oocyte and granulosa cells.Materials, Methods & Results: Thirty does were divided into three groups of 10 animals each, which received forage andconcentrate to provide respectively 2.7% of lipids (LL group), 3.9% lipids (LI group) and 5.1% lipids (LH group) for 28days. Three days before oocyte harvest, follicular wave was synchronized by 1 mL PGF2α intramuscularly, followed bythe insertion of an intravaginal progesterone release device. Viable oocytes and granulosa cells were subjected to qRT-PCRto determine the expression of PLIN2, ATF4, CHOP10, BAX, BCL2, HSP70 genes, were checked, evaluated using thedissociation curve analysis, which obtained the following values: 77.8, 80.2, 83.6, 78.0, 81.0 and 82.0ºC, respectively. Thesteps of qPCR thermal cycle was: denaturation and polymerase activation, annealing and final extension.Throughout theexperimental period, blood samples were taken for cholesterol, triglycerides and total lipids measurements. Total plasmalipids determination was calculated with the following equation: 2 x (cholesterol + triglycerides) × 1.1 with sensitivity ofthe assay for cholesterol was 1.472 mg/dL and for triglycerides, 2.845 mg/dL. In LH group, it was recorded a more pronounced increase in total lipids (+ 60.1%) (P < 0.05), cholesterol (+ 18.8%) and triglycerides (+ 8.5%). These increaseswere three times higher than that in LL and LI groups. All genes were expressed in oocytes...