Fidel Ovidio Castro- Ph.D
- Co-director Animal Biotechnology Lab at University of Concepción
Fidel Ovidio Castro
- Ph.D
- Co-director Animal Biotechnology Lab at University of Concepción
About
187
Publications
20,743
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Introduction
Main research projects: Stem cell research in farm animals; interested in optimizing ways to isolate MSC from cattle endometrium and to convert them into iPS cells. In horses and dogs, I am looking for molecules that can enhance the regenerative potential in tissue repair of adult MSC upon treatment of animals. Also I collaborate in a cattle cloning project and the study of mRNA and miRNA expression in cloned and normal cattle embryos at elongation
Current institution
Additional affiliations
December 2004 - October 2018
Education
November 2007 - March 2008
September 2001 - January 2002
September 1997 - May 1998
Klinik fur Tumor Biologie
Field of study
- Signal Transduction in Mammary Gland
Publications
Publications (187)
BACKGROUND
The physiopathology of life-threatening cerebrovascular complications in preeclampsia is unknown. We investigated whether disruption of the blood-brain barrier, generated using circulating small extracellular vesicles (sEVs) from women with preeclampsia or placentae cultured under hypoxic conditions, impairs the expression of tight junct...
Mare endometrosis is a major reproductive problem associated with low fertility and is characterized by persistent inflammation, TGFβ-1 signaling, and consequently, extracellular matrix deposition, which compromises endometrial glands. Mesenchymal stem cell-based products (MSCs), such as extracellular vesicles (EVs), have gained attention due to th...
Endometrosis in mares is a disease resulting from chronic inflammation characterized by peri glandular fibrosis. There is no effective treatment so far, which opens the door for exploring the use of stem cells as a candidate. Transforming growth factor beta (TGFβ) is crucial for the establishment and progression of fibrosis in mare´s endometrosis....
Background
Physiopathology of life-treating cerebrovascular complications in preeclampsia are yet unknown. We investigated whether disruption of the blood-brain barrier (BBB), generated using circulating small extracellular vesicles (sEVs) from women with preeclampsia or placentae cultured under hypoxic conditions, impairs the expression of tight j...
Domestic cat blastocysts cultured without the zona pellucida exhibit reduced implantation capacity. However, the protein expression profile has not been evaluated in these embryos. The objective of this study was to evaluate the protein expression profile of domestic cat blastocysts cultured without the zona pellucida. Two experimental groups were...
Simple Summary
In this study, it is described that embryos produced in vitro release nanoparticles charged with fragments of embryonic DNA. These nanoparticles, alongside their DNA content, can be recovered from the liquid medium where the embryos develop in vitro and used to assess the genetics of embryos. The analysis of gene markers is a common...
Domestic cat blastocysts cultured without the zona pellucida have a reduced implantation capacity and an altered expression of pluripotency and differentiation genes. Despite that, the protein expression profile has not been evaluated in these embryos. The objective of this study was to evaluate the protein expression profile of domestic cat blasto...
The modulation of inflammation is pivotal for uterine homeostasis. Here we evaluated the effect of the oestrus cycle on the expression of pro-inflammatory and anti-inflammatory markers in a cellular model of induced fibrosis. Mare endometrial stromal cells isolated from follicular or mid-luteal phase were primed with 10 ng/mL of TGFβ alone or in co...
The in vitro culture of domestic cat embryos without the zona pellucida affects their implantation capacity. MicroRNAs (miRNAs) have an important role in embryo–maternal communication and implantation. The objective of this study was to evaluate the expression of specific miRNAs in domestic cat blastocysts cultured without the zona pellucida. Two e...
Background. The domestic cat is a valuable model for the development of assisted
reproductive technologies in endangered felids. However, domestic cat embryos cultured without the zona pellucida are not able to implant (1). The reason for this lack of implantation is still unknown. The objective of this study was to evaluate the expression of speci...
The embryo-maternal interaction occurs during the early stages of embryo development and is essential for the implantation and full-term development of the embryo. In bovines, the secretion of interferon Tau (IFNT) during elongation is the main signal for pregnancy recognition, but its expression starts around the blastocyst stage. Embryos release...
Pre-implantation embryos release extracellular vesicles (EVs) to extracellular environment. In this work it is hypothesized that the EVs miRNA cargo will vary during pre-implantation development due to the constant changes in gene expression that take place through this period. The concentration, size and miRNA cargo of EVs secreted by competent bo...
Domestic cat blastocysts cultured without zona pellucida have a reduced implantation capacity after embryo transfer. The objective of this study was to evaluate the expression pattern of trophectoderm markers in domestic cat blastocysts cultured without zona pellucida. For this, two experimental groups were evaluated: (1) Domestic cat embryos gener...
Domestic cat embryos generated by in vitro fertilization (IVF) and cultured without the zona pellucida have a reduced implantation capacity after embryo transfer at the blastocyst stage. The objective of this study was to evaluate the expression of trophectoderm markers in domestic cat blastocysts cultured without the zona pellucida. Two experiment...
BACKGROUND-AIM Domestic cat embryos cultured without zona pellucida have a reduced implantation capacity after embryo transfer at the blastocyst stage. The objective of this study was to evaluate the expression of trophectoderm markers in domestic cat blastocysts cultured without zona pellucida. METHODS Two experimental groups were done: 1) Domesti...
BACKGROUND-AIM
In dairy cattle, follicular development and oocyte quality depends on age, production performance, reproductive status and other factors affecting the success of ovum pick
up (OPU) and subsequently in vitro embryo production, thus it is mandatory to standardize protocols for oocyte recovery considering particular characteristics of d...
BACKGROUND-AIM
Although the oocyte collection by ovum pick up (OPU) is performed with minimal stress to the donor the repeated OPU practice may influence cattle productivity. The present
study evaluated the effects of repeated OPU practice on oocyte recovery rates (ORRs) and milk production performance in dairy cows of different parity status.
METH...
This work aims to evaluate the efficiency of detecting embryonic DNA in extracellular vesicles (EVs) secreted by in vitro produced bovine embryos. It is hypothesized that embryonic DNA is released within EVs, and thus, the isolation of these vesicles would improve the use of spent medium as a noninvasive method for embryo sexing and genotyping. The...
Over the last few years, several commercial FSH products have been developed for cattle superovulation (SOV) purposes in Multiple Ovulation and Embryo Transfer (MOET) programs. The SOV response is highly variable among individuals and remains one of the main limiting factors in obtaining a profitable number of transferable embryos. In this study, f...
The domestic cat is a valuable model for development of assisted reproductive techniques in endangered felids. Roscovitine (RO) is a reversible inhibitor of M-phase promoting factor (MPF) that generates a delay in maturation, thus improving oocyte competence. However, prematuration with RO negatively affects the viability of cumulus cells and in vi...
Preeclampsia, a pregnancy-related endothelial disorder, is associated with both cardiovascular and cerebrovascular complications. Preeclampsia requires the presence of a placenta as part of its pathophysiology, yet the role of this organ in the cerebrovascular complications remains unclear. Research has shown that circulating small extracellular ve...
Roscovitine (RO) is a reversible inhibitor of MPF. Incubation of immature COCs with RO followed by IVM enhances oocyte competence. Additionally, it is know that SCNT is a complex procedure in which only a limited number of viable embryos can be generated. The reversible delay on oocyte maturation generated by RO might enhances the cytoplasmic matur...
The removal of the zona pellucida has been used to improve the in vitro development of domestic cat embryos generated by IVF and SCNT. However, the in vivo development of domestic cat embryos generated without the zona pellucida has not been evaluated. The objective of this study was to evaluate the effects of zona pellucida removal on the in vitro...
As células-tronco mesenquimais (CTM) são utilizadas na terapia celular, isolando-se de diferentes tecidos, incluindo a pele. Os fibroblastos dérmicos têm mostrado características de potência semelhantes às CTMs como expressão de marcadores de superfície e diferenciação para outros tipos de linhagens de origem mesodérmica, principalmente sob a influ...
Extracellular vesicles are nanoparticles secreted by cell and have been proposed as suitable markers to identify competent embryos produced in vitro. Characterizing EVs secreted by individual embryos is challenging because culture medium itself contributes to the pool of nanoparticles that are co-isolated. To avoid this, culture medium must be depl...
Routine blood analysis indicated the presence of Mycoplasma-like bodies in a guigna (Leopardus guigna). Evidence of infection with Candidatus Mycoplasma haemominutum was found in blood samples using PCR and DNA sequencing of the 16S rRNA gene of Mycoplasma. Mycoplasma spp. are documented in cats but their role in the transmission of Mycoplasma to g...
During early development, embryos secrete extracellular vesicles (EVs) that participate in embryo–maternal communication. Among other molecules, EVs carry microRNAs (miRNAs) that interfere with gene expression in target cells; miRNAs participate in embryo–maternal communication. Embryo selection based on secreted miRNAs may have an impact on bovine...
Human embryos generated in vitro have a high incidence of chromosomal abnormalities that negatively affect pregnancy rate. Embryos generated in vitro secrete extracellular vesicles (EVs) into the culture medium that could be used potentially as indicators of embryo competence. This research aimed to evaluate the concentration and size of EVs and th...
Equine endometrial and adipose mesenchymal stem cells (eMSCs and aMSCs, respectively) were isolated from the same donors of thoroughbred mares. The cells displayed characteristic features of MSCs, including trilineage mesodermal and also neurogenic differentiation. We evaluated the influence of cellular origin on their transcriptome profile. Cellul...
Estrogenic steroids and adenosine A2A receptors promote the wound healing and angiogenesis processes. However, so far, it is unclear whether estrogen may regulate the expression and pro-angiogenic activity of A2A receptors. Using in vivo analyses, we showed that female wild type (WT) mice have a more rapid wound healing process than female or male...
Abstract
The kodkod (Leopardus guigna) is a small felid endemic of Chile and is considered a vulnerable species. Domestic cat oocytes have been successfully used as recipient cytoplast to reprogram somatic cells from different felids by interspecific somatic cell nuclear transfer (iSCNT). The developmental competence of felid embryos generated by i...
Mesenchymal stem cells (MSC) are modern tools in regenerative therapies of humans and animals owed to their immunomodulatory properties, which are activated in a pro-inflammatory environment. Different preconditioning strategies had been devised to enhance the immunomodulatory properties of MSC. In this research, we evaluated the immunological attr...
Simple Summary
Prostaglandin E2 plays a regulatory role in MSC self-renewal, and PGE2 receptor-mediated signalling is involved in cell proliferation and migration. The aim of this work was to establish a genetically modified equine cellular model in which receptor genes EP2 and EP4 were edited by the CRISPR/Cas9 system. We successfully targeted EP2...
Adipose derived mesenchymal stem cells (AMSCs) have been isolated from domestic and wild cats. For wild cats, the isolation of AMSCs has been reported in the black-footed cats (Felis nigripes) and guigna (Leopardus guigna). Stromal vascular fraction (SVF) isolated from cougar adipose tissue have been used to restore elbow functionality in the couga...
The objective of this study was to evaluate, in the domestic cat, the effect of ovarian stimulation with eCG prior to oocyte in vitro maturation (priming) on in vitro and in vivo development after in vitro fertilization (IVF). For this purpose, oocyte donors were either 1) treated with a single dose of 200 IU eCG four days before oocyte recovery (e...
Background: Endometrosis is a multifactorial disease and one of the main causes of infertility in mares, its etiology and pathogenesis are not completely understood. It is defined as peri glandular and/or stromal endometrial fibrosis with glandular alterations. Due to the few clinical symptoms, besides anamnesis and fertility data, endometrosis req...
Noninvasive methods are the clue to increase the efficiency of invitro-derived embryo selection without decreasing their competence. Embryos selection based on their morphology is the most used method but only 40% of selected embryos are able to implant and develop correctly. In humans, pre-implantation genetic diagnosis increases the efficiency of...
The success of development of invitro embryo production needs to mimic culture conditions in the maternal environment. Recently, it has been seen that extracellular vesicles (EVs) secreted by oviducal or endometrial cells may improve development and quality of embryos produced invitro. Extracellular vesicles are a mechanism of cellular communicatio...
Post-mating induced endometritis (PMIE) is an acute inflammatory response of the endometrium to spermatozoa, linked to an incapability of some mares to drain out the fluids associated with inflammation. This is of pivotal importance for reproductive success in mares. Mesenchymal stem cells (MSCs) are potential candidates for anti-inflammatory uteri...
Reprogramming of terminally differentiated cells to higher plasticity levels can be achieved with small molecules. This can be of value for somatic cell nucleus transfer, deriving multi and pluripotent cells and conservation purposes. Recently, induced mesenchymal stem cells were derived from differentiated human and mouse cells by using small mole...
Extracellular vesicles (EVs), including exosomes and microvesicles, are secreted by different cell types and participate in cellular communication by carrying molecules as microRNAs (miRNAs) that can interfere with gene expression of target cells. Extracellular vesicles have become relevant as a mechanism of embryo-maternal communication. The aim o...
Limited information have been reported regarding to the role of the zona pellucida during embryo development in the domestic cat. In the present research we compared invitro and invivo development of cat embryos generated with versus without the zona pellucida. Embryos were produced by IVF and cultured up to blastocyst stage accordingly to experime...
Adipose derived mesenchymal stem cells (AMSCs) have been isolated from domestic and wild cats. For wild cats, the isolation of AMSCs has been reported in the black-footed cats (Felis nigripes) and guigna (Leopardus guigna). Stromal vascular fraction (SVF) isolated from cougar adipose tissue have been used to restore elbow functionality in the couga...
Quality of in vitro -produced embryos is influenced by changes in gene expression in response to adverse conditions. Gene markers for predicting ‘good embryos’ do not exist at present. We propose that the expression of pluripotency markers OCT4 – SOX2 – NANOG in D9 (day 9) bovine demi-embryos correlated with development at D13 (day 13). Day 8 in vi...
Extracellular vesicles (EVs) secreted by blastocysts may be clinically relevant, as indicator of embryo viability on in vitro fertilization. We tested if the characteristics of extracellular vesicles secreted during blastulation are related to embryo viability. Morulae were individually cultured in SOF media depleted of EVs until day 7.5 post IVF....
The domestic cat is a valuable model for the development of assisted reproductive techniques that might be used in the conservation of endangered felids. However, the efficiency of the in vitro embryo production in the domestic cat remains low compared to other species. In the bovine and equine, the zona pellucida removal enhance the developmental...
Adipose tissue derived mesenchymal stem cells (AMSCs) had been isolated and used for cell therapy in domestic cats. For wild cats, the isolation of AMSCs has only been reported in the black-footed cat (Felis nigripes). AMSCs obtained from wild cats may be useful to treat injuries of endangered cat species that remain in captivity or arrive at wildl...
Earlier we successfully isolated and characterised endometrial (eMSC) and adipose (aMSC) mesenchymal stem cells from the same donors. Mesenchymal stem cells share biological traits but display different surface marker phenotype and migration ability. Here we extended our research to their mRNA signature using next-generation sequencing. The RNA fro...
Prostaglandin E2 (PGE2) acts through 4 cellular receptors: EP1, EP2, EP3, and EP4; only EP2 and EP4 are relevant for immunomodulation and migration in immune cells. Besides those, several cells express these receptors on their surface, including mesenchymal stem cells. Pharmacological inhibition of the EP2 receptor prevents migration of immune syst...
The kodkod is considered a vulnerable species by the International Union for Conservation of Nature. Phylogenetically, the kodkod is classified in the Leopardus genus, which has only 36 chromosome pairs compared with the domestic cat, which has 38. The proposed hypothesis was that domestic cat oocytes are capable of reprogramming somatic cells from...
Extracellular vesicles (EV) secreted by blastocysts might be relevant to predict competence of embryos produced in vitro. The aim of this study was to develop a model to select competent embryos that combines blastocyst morphokinetics data and morphological parameters of EV secreted during blastulation (Days 5-7.5). Embryos were cultured in groups...
Bovine embryos that rapidly reach the two-cell stage have higher probabilities of becoming viable blastocysts. Our objective was to evaluate the effect of time of the first embryonic division on the development rate of bovine blastocysts produced in vitro and also to determine embryo viability by differential staining of living and dead blastomeres...
The high metabolic activity to which the dairy cattle are exposed for maintain milk production altered steroid metabolism that affect reproductive physiology and reducing oocyte competence. Our aims were 1) to characterize the competence of immature oocytes collected from dairy cattle based on the expression of genes in cumulus cells and 2) to impr...
El uso de células madre mesenquimales (MSCs) aisladas de varios tejidos representan una oportunidad para ser empleadas en terapia celular en animales domésticos y silvestres. Aunque en gatos domésticos se ha descrito ampliamente su aislamiento y caracterización desde diferentes tejidos, en felinos salvajes existen pocos registros de aislamiento de...
Obtaining mesenchymal stem cells (MSC) might be troublesome for some species, particularly the feline. Further, in patients with complex pathologies or trauma like wild cats, when biopsy sampling is an issue, it can be even more complicated. The objective of this study was to produce h-PDGF-B by stable transfection of human SiHa cells for future us...
En la actualidad, se describe un aumento en los trabajos de investigación relacionados con el desarrollo de técnicas de reproducción asistida en el gato doméstico. Esto con el propósito de implementar posteriormente estas técnicas en la conservación de felinos silvestres amenazados (Pope, 2014). Sin embargo, los sistemas de producción in vitro de e...
The aim of this research was to characterize at the mRNA level, the response of bovine endometrial mesenchymal stem cells (beMSCs) to an inflammatory environment after priming with PGE2. For this, we used previously tested beMSC lines from late luteal phase of the oestral cycle, which showed an altered expression of immune response genes after PGE2...
Here we aimed to isolate and characterize bovine endometrial mesenchymal stem cells(beMSCs)during follicular(FP),early luteal(ELP)and late luteal(LLP)phases of the estral cycle in healthy cows and after parturition in healthy(PPHE)and in cows with subclinical or clinical endometritis(PPSEandPPCE).The estral cycle stages were confirmed by measuring...
The endometrium is an accessible source of mesenchymal stem cells. Most investigations of endometrial mesenchymal stem cells (eMSCs) have been conducted in humans. In animals, particularly in livestock, eMSC research is scarce. Such cells have been described in the bovine, ovine, caprine, porcine, and equine endometrium. Here we provide the state o...
Contents
The objective of this study was to evaluate the eCG stimulation on domestic cat oocyte competence during the non‐breeding season. Four experimental groups were made: (a) untreated cycling cats (Breeding season group; BS), (b) untreated anestrous cats (Non‐breeding group; NB), (c) anestrous cats treated with 200 IU of eCG (eCG group) and (d...
Cell therapy is an option currently used for treatment of several pathologies in many species. Cat adipose mesenchymal stem cells (MSCs) have been used successfully in certain therapies in domestic cats. However, availability and easiness of access to these cells in feline patients has been an issue. Mouse embryonic fibroblast as well as terminally...
Questions
Questions (5)
Hi all, here in Chile we are facing a brutal shortage of consumables of Merck-Sigma-Millipore, specially after the fusion of Merck with Sigma and the pandemia, anyway the thing is that ultra filters from Millipore are taking 6 to 8 months to reach us! and we base many of our projects in ultrafiltration. The question es: do you know of a reliable (trsutable) substitutes for Amicon ultrafilters? Gracias
Hi all, many years ago I used a plasmid for over expressing an ovine mammary gland factor (later known as STAT5a). It was named pXM-oMGF, I do have the plasmid in my hands, but can not get the map of it, neither its antibiotic resistance (for pro and eukaryotes). I did my best in the literature and followed the thread, but unfortunately none of the papers described the actual plasmid, just mention that it was described somewhere else and when I get there, it sends me to another and so on. Also can not find in Addgene, or other databases, the pXM family from which must likely stemmed to see the resistance. I do not have my laboratory notebook of those years any longer and really need the information. If any body can help please?!!! Thank you
Hi there, I need to perform dot or slot blot for proteins, but I do not have a manifold device. I recall when I was "younger" people use to dry out membranes without vacuum. Any suggestion?
Hi, I need to quantify the levels of cAMP intracellularly in a cell line after stimulation of the EP2 receptor with PGE2. In the web I found severa protocols, most of them using radioactivity, which I can not use in my lab. I would appreciate any advice. Thanks Fidel
Particularly I am concern about using human antibodies that might not recognize the horse antigens and also not sure if labelling my primary antibodies will actually do the job?
