185 EMBRYONIC LOSS IN PIGS ASSOCIATED WITH OVIDUCT TRANSPLANTATION OF EARLY-STAGE EMBRYOS WITH DAMAGES IN THE ZONA PELLUCIDA

2006 ◽  
Vol 18 (2) ◽  
pp. 200
Author(s):  
S. Ueno ◽  
M. Kurome ◽  
R. Tomii ◽  
K. Hiruma ◽  
N. Maeda ◽  
...  
Keyword(s):  
Immune Response ◽  
Zona Pellucida ◽  
Nuclear Transfer ◽  
Early Stage ◽  
Giant Cells ◽  
Blastocyst Stage ◽  
Embryo Recovery ◽  
Embryonic Loss ◽  
The Impact

It is assumed that if porcine early-stage embryos with damages in their zonae pellucidae are transplanted to the recipient's oviduct, they may suffer from mechanical and immunological stresses by oviduct contraction and the recipient's immune response. This study aimed to examine the impact of zona pellucida damages, which might arise during nuclear transfer and intra cytoplasmic sperm injection (ICSI), on the development and survival of transplanted embryos. Cumulus-oocyte complexes were collected from ovaries obtained at a local slaughterhouse and matured in vitro in NCSU23 to prepare MII-stage oocytes. The zonae pellucidae of these oocytes were either penetrated with 8- to 10-�m square-ended microinjection pipettes or incised with 35- to 40-�m beveled enucleation pipettes. Intact oocytes were used as controls. The oocytes were electroactivated to induce parthenogenesis and transplanted to the oviducts of estrus-synchronized recipient gilts (estrus-synchronized with 1000 IU eCG and 1500 IU hCG). After 5 to 7 days, the recipient uteri were flushed with PBS supplemented with 1% fetal bovine serum (FBS) to collect embryos, and their development (morula-blastocyst stage embryos/collected embryos) and survival (viable embryos/collected embryos) were determined. In total, 221 zona-penetrated, 129 zona-incised, and 57 intact embryos were transplanted to four, two and two gilts, respectively. The efficiency of embryo recovery was similar in all groups (59.0 to 81.8%). However, the zona-penetrated and zona-incised embryos showed inconsistent development and survival compared with controls; the development and survival rate were 92.6% (25/27) to 96.7% (29/30) and 77.8% (21/27) to 96.7% (29/30) in control embryos, respectively, whereas those of zona-penetrated embryos were 57.1% (28/49) to 95.7% (22/23) and 8.2% (4/49) to 78.3% (18/30), and those of zona-incised embryos were 47.6% (30/63) to 92.3% (36/39) and 23.8% (15/63) to 92.3% (22/23), respectively. Large foci of cells that appeared to be macrophage giant cells were observed at the surface or inside of the degenerated zona-damaged embryos. These results indicate that the recipient's immune response may impair development after transplantation of the embryo to the oviduct, when there is damage in the zona pellucida. This may be one of the factors attributable to the reduced efficiency of live progeny production by ICSI and nuclear transfer. This work was supported by PROBRAIN.

scholarly journals Paraquat Preferentially Induces Apoptosis of Late Stage Effector Lymphocyte and Impairs Memory Immune Response in Mice

2019 ◽  
Vol 16 (11) ◽  
pp. 2060 ◽  
Author(s):  
Yiming Shao ◽  
Yifan Zhao ◽  
Tingting Zhu ◽  
Fen Zhang ◽  
Xiuli Chang ◽  
...  
Keyword(s):  
Immune Response ◽  
Late Stage ◽  
Early Stage ◽  
Keyhole Limpet Hemocyanin ◽  
Primary Immune Response ◽  
Secondary Immune Response ◽  
Effector Cells ◽  
Memory Immune Response ◽  
The Impact

Paraquat (PQ) is a toxic non-selective herbicide. To date, the effect of PQ on memory immune response is still unknown. We investigated the impact of PQ on memory immune response. Adult C57BL/6 mice were subcutaneously injected with 2 mg/kg PQ, 20 mg/kg PQ or vehicle control every three days for two weeks. A single injection of keyhole limpet hemocyanin (KLH) at day four after the initial PQ treatment was used to induce a primary immune response; a second KLH challenge was performed at three months post the first KLH immunization to induce a secondary immune response. In steady state, treatment with 20 mg/kg PQ reduced the level of serum total IgG, but not that of IgM; treatment with 20 mg/kg PQ decreased the number of effector and memory lymphocytes, but not naïve or inactivated lymphocytes. During the primary immune response to KLH, treatment with 20 mg/kg PQ did not influence the proliferation of lymphocytes or expression of co-stimulatory molecules. Instead, treatment with 20 mg/kg PQ increased the apoptosis of lymphocytes at late stage, but not early stage of the primary immune response. During the secondary immune response to KLH, treatment with 20 mg/kg PQ reduced the serum anti-KLH IgG and KLH-responsive CD4 T cells and B cells. Moreover, effector or activated lymphocytes were more sensitive to PQ-induced apoptosis in vitro. Treatment with 2 mg/kg PQ did not impact memory immune response to KLH. Thus, treatment with 20 mg/kg PQ increased apoptosis of late stage effector cells to yield less memory cells and thereafter impair memory immune response, providing a novel understanding of the immunotoxicity of PQ.

Early embryo development in nuclear transfer clones derived from somatic cells of a high genetic merit dairy cow

2001 ◽  
Vol 26 (2) ◽  
pp. 437-440
Author(s):  
W.H. McMillan ◽  
D.N. Wells ◽  
AJ. Peterson ◽  
M.J. Donnison
Keyword(s):  
Embryo Transfer ◽  
Nuclear Transfer ◽  
Early Stage ◽  
Somatic Cells ◽  
Early Embryo ◽  
Embryo Morphology ◽  
Embryo Recovery ◽  
Stage Of Development ◽  
Developmental Traits

AbstractThe reported post transfer survival rate to term of cloned embryos derived from both undifferentiated blastomeres and differentiated foetal and adult cells is very low (typically less than 20%). Furthermore, it is acknowledged that there are many technical issues that remain to be resolved to improve the efficiency of nuclear transfer before the technique will find widespread, practical and cost-effective use in multiplying valuable livestock in agriculture. The purpose of this study was to compare early embryo morphology following embryo transfer of nuclear transfer blastocysts derived from somatic cells of an adult Friesian cow with that of standard in vitro-produced embryos. In the present study, 150 embryos were transferred in bulk (i.e., 15, 20 or 25 per recipient) to the ipsilateral uterine horn of 8 recipients using standard non-surgical embryo transfer procedures. Embryos were then recovered following necropsy on either Day 14 or Day 23 of pregnancy and developmental traits described. Embryo recovery and elongation rates were similar on Day 14 of pregnancy (Table 1), although cloned conceptuses were longer and narrower (P<0.05). Embryo recovery and viability rates by Day 23 were similar, although many of the developmental traits appeared more advanced in cloned conceptuses. (Table 1). Allantois development was different because of greater widths and the presence of ‘spurs’ that were not observed with in vitro-produced embryos. We conclude that apparently abnormal conceptus development occurs by about 3 weeks of pregnancy following nuclear transfer, but that embryo survival is not compromised at this early stage of development.

Embryo manipulation in cattle breeding and production

1989 ◽  
Vol 48 (1) ◽  
pp. 3-30 ◽  
Author(s):  
J. A. Woolliams ◽  
I. Wilmut
Keyword(s):  
Gene Transfer ◽  
Beef Cattle ◽  
Embryo Transfer ◽  
Nuclear Transfer ◽  
Progeny Testing ◽  
Genetic Progress ◽  
Embryo Recovery ◽  
Multiple Ovulation ◽  
The Impact

ABSTRACTDevelopments, both recent and potential, in procedures for manipulating embryos are described. The procedures considered include: embryo transfer, multiple ovulation and embryo recovery, recovery of oocytes, in vitro maturation (IVM) and fertilization (IVF) of oocytes, in vitro culture of zygotes, embryo splitting and nuclear transfer, embryo storage, embryo sexing, gene transfer and embryo stem cells. The impact of these procedures on breeding strategies such as multiple ovulation and embryo transfer (MOET) nucleus breeding schemes and progeny testing are discussed for both dairy and beef cattle.For MOET nucleus schemes all these procedures have potential applications in producing maximal rates of genetic progress for a fixed rate of inbreeding. With the current effectiveness of the procedures, embryo sexing and nuclear transfer would have the most impact. The potential for increasing genetic progress through progeny testing is enhanced using multiple ovulation, embryo recovery and transfer in cows to breed bulls, but no other procedures appeared to offer major benefits. The efficiency of beef production from the dairy herd could be increased either by using IVM and IVF to produce more beef-type calves or, potentially, by cloning and embryo transfer, to produce pure beef calves. Procedures leading to the production of clone families would make an impact on the evaluation of genotypes and environments. Gene transfer may be used to modify the composition of milk including the production of pharmaceutical proteins, and to increase milk yield or the efficiency of lean meat production.It is concluded that, although much further research is required, the procedures discussed will have major implications for the structure and organization of dairy and beef cattle herds over the next decade.

scholarly journals Developmental disparity between in vitro-produced and somatic cell nuclear transfer bovine days 14 and 21 embryos: implications for embryonic loss

Reproduction ◽  
2008 ◽  
Vol 136 (4) ◽  
pp. 433-445 ◽  
Author(s):  
Natalie I Alexopoulos ◽  
Poul Maddox-Hyttel ◽  
Pernille Tveden-Nyborg ◽  
Nancy T D'Cruz ◽  
Tayfur R Tecirlioglu ◽  
...  
Keyword(s):  
Somatic Cell ◽  
Somatic Cell Nuclear Transfer ◽  
Nuclear Transfer ◽  
Reproductive Technologies ◽  
Embryonic Mortality ◽  
High Rate ◽  
Morphological Examination ◽  
Embryonic Loss ◽  
The Impact

In ruminants, the greatest period of embryonic loss coincides with the period of elongation when the embryonic disc is formed and gastrulation occurs prior to implantation. The impact of early embryonic mortality is not only a major obstacle to the cattle breeding industry but also impedes the application of new reproductive technologies such as somatic cell nuclear transfer (SCNT). In the present study, days 14 and 21 bovine embryos, generated by eitherin vitro-production (IVP) or SCNT, performed by either subzonal injection (SUZI) or handmade cloning (HMC), were compared by stereomicroscopy, immunohistochemistry, and transmission electron microscopy to establishin vivodevelopmental milestones. Following morphological examination, samples were characterized for the presence of epiblast (POU5F1), mesoderm (VIM), and neuroectoderm (TUBB3). On D14, only 25, 15, and 7% of IVP, SUZI, and HMC embryos were recovered from the embryos transferred respectively, and similar low recovery rates were noted on D21, suggesting that most of the embryonic loss had already occurred by D14. A number of D14 IVP, SUZI, and HMC embryos lacked an epiblast, but presented trophectoderm and hypoblast. When the epiblast was present, POU5F1 staining was limited to this compartment in all types of embryos. At the ultrastructural level, SCNT embryos displayed abundant secondary lysosomes and vacuoles, had fewer mitochondria, polyribosomes, tight junctions, desmosomes, and tonofilaments than their IVP counterparts. The staining of VIM and TUBB3 was less distinct in SCNT embryos when compared with IVP embryos, indicating slower or compromised development. In conclusion, SCNT and to some degree, IVP embryos displayed a high rate of embryonic mortality before D14 and surviving embryos displayed reduced quality with respect to ultrastructural features and differentiation markers.

80 EFFECT OF MEDIA, SYNCHRONIZATION OF FIBROBLAST CELLS, CULTURE TIME, OXYGEN CONCENTRATION, AND ACTIVATION ON NUCLEAR-TRANSFERRED PORCINE OOCYTES

2007 ◽  
Vol 19 (1) ◽  
pp. 157
Author(s):  
J. H. Quan ◽  
H. B. Seok ◽  
S. K. Kim
Keyword(s):  
Nuclear Transfer ◽  
Culture Medium ◽  
Atmospheric Condition ◽  
Blastocyst Stage ◽  
Fibroblast Cells ◽  
Fetal Fibroblast ◽  
Donor Cells ◽  
The Impact ◽  
Cells Cultured

The purpose of this study was to investigate the impact of culture medium, culture duration, and atmospheric condition on the fusion and in vitro development rates of nuclear transfer porcine embryos constructed by the microinjection of fetal fibroblast cells into in vitro-matured oocytes. Single fetal donor cells were deposited into the perivitelline space of enucleated oocytes, followed by electrical fusion and activation. Activated embryos were cultured in NCSU-23 medium supplemented with 5% FBS, at 38.5�C for 6 to 8 days in 5% CO2 and air. In Experiment 1, the fusion rates of nuclear transfer embryos did not differ from those of fetal fibroblast cells incubated in 5% FBS + NCSU-23 or 5% FBS + TL-HEPES medium, nor did fusion rates of donor cells differ among 1–8-h incubation durations. Fusion rates for the 4 treatment subclasses ranged from 72.1% to 78.0%. In Experiment 2, pre-synchronization in medium containing 0.1 �g mL-1 Hoechst(H) 33342 increased during the period from 0 and 8 h of culture up to 15 h, the end of the synchronization period, at which time there was a significantly increased percentage of porcine fibroblast cells at the G2/M stage (12.4%, 17.5%, and 47.6%; P &lt; 0.01). Neither an increase in the concentration of H 33342 (0.2–1.6 �g mL-1) nor a longer exposure time (12 h, 18 h, and 24 h) increased the proportion of porcine G2/M fibroblasts. In Experiment 3, fusion rates did not differ significantly between nuclear transfer embryos constructed using donor cells cultured in 5% FBS + NCSU-23 medium for 1–2, 6–8, or 12–14 days (60.0%, 73.3%, and 62.5%, respectively). The cleavage rate for nuclear transplant embryos using fetal fibroblast cells cultured for 1–2 days was 44.0%, which was significantly less than the 56.7% and 50.0% for 6–8 or 12–14 days of culture, respectively (P &lt; 0.05). In Experiment 4, the proportions of nuclear transfer embryos that developed to the e2 cell and to the blastocyst stage were not affected significantly by culture medium (5% FBS + NCSU-23 or 5% FBS + TL-HEPES) or by O2 concentration during culture (5% vs. 10%). The developmental rates to the e2 cell stage ranged from 65.9% to 70.1%, and those to the blastocyst stage ranged from 9.8% to 12.5%, for the 4 treatment subclasses. Blastocyst rate was highest for embryos cultured in 5% FBS + NCSU-23 under a gas atmosphere of 5% O2 in air.

84 PRODUCTION OF CLONED PIGLETS FROM NUCLEAR TRANSFER EMBRYOS AFTER VITRIFICATION

2008 ◽  
Vol 20 (1) ◽  
pp. 123
Author(s):  
N. Nakayama ◽  
R. Tomii ◽  
S. Ueno ◽  
H. Matsunari ◽  
H. Saito ◽  
...  
Keyword(s):  
Zona Pellucida ◽  
Nuclear Transfer ◽  
Lipid Droplets ◽  
Cytochalasin B ◽  
Calf Serum ◽  
Blastocyst Stage ◽  
Early Cleavage ◽  
C 23 ◽  
Morula Stage

Cryopreservation of cloned embryos is expected to be beneficial in improving the efficiency of somatic cell cloning in pigs. We have already demonstrated that normal piglets can be produced from in vitro-matured and fertilized (IVM/IVF) embryos vitrified at an early cleavage stage after delipation (Nagashima et al. 2007 Biol. Reprod. 76, 900–905). In this study we utilized this technique in an attempt to produce piglets from cloned embryos reconstructed with IVM oocytes. Nuclear transfer (NT) embryos were reconstructed using oocytes matured in vitro in NCSU23 and preadipocytes as nuclear donors. The embryos were cultured in PZM-5 for approximately 98 h, and those that had developed to the morula stage were delipated using a noninvasive method described previously (Esaki et al. 2004 Biol. Reprod. 71, 432–437). The embryos were treated with 4% trypsin at 38�C for 1 to 4 min to induce a slight swelling of the zona pellucida, and then centrifuged (12 000g, 38�C, 23 min) with 7.5 µg mL–1 cytochalasin B to polarize cytoplasmic lipid droplets within the perivitelline space. The embryos were cultured for 1 h and vitrified by the minimum volume cooling (MVC) method using a MVC plate (Cryotop�; Kitasato Supply Co., Tokyo, Japan) in the presence of 15% ethylene glycol, 15% DMSO, and 0.5 m sucrose as cryoprotectants. Vitrified embryos were rewarmed by immersing the MVC plate diretly into rewarming solution containing 1 m sucrose and 20% calf serum at 38�C for 1 min, followed by stepwise dilution of the cryoprotectants. The rewarmed embryos were cultured for 2 days to the blastocyst stage, and then treated with 0.5% pronase to remove the zona pellucida before transfer to the uterine horn of recipients. A total of 103 vitrified blastocysts were transferred to 2 recipient gilts. Both gilts became pregnant and farrowed 2 and 4 piglets, respectively (6/103, 5.8%). These results demonstrate that cloned piglets can be produced from NT embryos that have been cryopreserved at the morula stage using noninvasive delipation and vitrification procedures. This study was supported by PROBRAIN.

scholarly journals Decitabine Promotes Modulation in Phenotype and Function of Monocytes and Macrophages That Drive Immune Response Regulation

Cells ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 868
Author(s):  
Fabiana Albani Zambuzi ◽  
Priscilla Mariane Cardoso-Silva ◽  
Ricardo Cardoso Castro ◽  
Caroline Fontanari ◽  
Flavio da Silva Emery ◽  
...  
Keyword(s):  
Immune Response ◽  
Hematological Malignancies ◽  
M2 Macrophage ◽  
M2 Polarization ◽  
Tnf Α ◽  
Monocytes And Macrophages ◽  
Ifn Γ ◽  
And Function ◽  
The Impact

Decitabine is an approved hypomethylating agent used for treating hematological malignancies. Although decitabine targets altered cells, epidrugs can trigger immunomodulatory effects, reinforcing the hypothesis of immunoregulation in treated patients. We therefore aimed to evaluate the impact of decitabine treatment on the phenotype and functions of monocytes and macrophages, which are pivotal cells of the innate immunity system. In vitro decitabine administration increased bacterial phagocytosis and IL-8 release, but impaired microbicidal activity of monocytes. In addition, during monocyte-to-macrophage differentiation, treatment promoted the M2-like profile, with increased expression of CD206 and ALOX15. Macrophages also demonstrated reduced infection control when exposed to Mycobacterium tuberculosis in vitro. However, cytokine production remained unchanged, indicating an atypical M2 macrophage. Furthermore, when macrophages were cocultured with lymphocytes, decitabine induced a reduction in the release of inflammatory cytokines such as IL-1β, TNF-α, and IFN-γ, maintaining IL-10 production, suggesting that decitabine could potentialize M2 polarization and might be considered as a therapeutic against the exacerbated immune response.

scholarly journals NLRP7 Promotes Choriocarcinoma Growth and Progression through the Establishment of an Immunosuppressive Microenvironment

Cancers ◽  
2021 ◽  
Vol 13 (12) ◽  
pp. 2999
Author(s):  
Deborah Reynaud ◽  
Roland Abi Nahed ◽  
Nicolas Lemaitre ◽  
Pierre-Adrien Bolze ◽  
Wael Traboulsi ◽  
...  
Keyword(s):  
Immune Response ◽  
Tumor Cells ◽  
Major Gene ◽  
Critical Role ◽  
Orthotopic Model ◽  
Independent Manner ◽  
Maternal Immune Response ◽  
The Impact

The inflammatory gene NLRP7 is the major gene responsible for recurrent complete hydatidiform moles (CHM), an abnormal pregnancy that can develop into gestational choriocarcinoma (CC). However, the role of NLRP7 in the development and immune tolerance of CC has not been investigated. Three approaches were employed to define the role of NLRP7 in CC development: (i) a clinical study that analyzed human placenta and sera collected from women with normal pregnancies, CHM or CC; (ii) an in vitro study that investigated the impact of NLRP7 knockdown on tumor growth and organization; and (iii) an in vivo study that used two CC mouse models, including an orthotopic model. NLRP7 and circulating inflammatory cytokines were upregulated in tumor cells and in CHM and CC. In tumor cells, NLRP7 functions in an inflammasome-independent manner and promoted their proliferation and 3D organization. Gravid mice placentas injected with CC cells invalidated for NLRP7, exhibited higher maternal immune response, developed smaller tumors, and displayed less metastases. Our data characterized the critical role of NLRP7 in CC and provided evidence of its contribution to the development of an immunosuppressive maternal microenvironment that not only downregulates the maternal immune response but also fosters the growth and progression of CC.

scholarly journals Miniaturization and Automation of a Human In Vitro Blood–Brain Barrier Model for the High-Throughput Screening of Compounds in the Early Stage of Drug Discovery

Pharmaceutics ◽  
2021 ◽  
Vol 13 (6) ◽  
pp. 892
Author(s):  
Elisa L. J. Moya ◽  
Elodie Vandenhaute ◽  
Eleonora Rizzi ◽  
Marie-Christine Boucau ◽  
Johan Hachani ◽  
...  
Keyword(s):  
Drug Discovery ◽  
Blood Brain Barrier ◽  
Early Stage ◽  
Molecular Transport ◽  
Brain Barrier ◽  
Blood Brain ◽  
Cns Drugs ◽  
The Impact ◽  
The Brain

Central nervous system (CNS) diseases are one of the top causes of death worldwide. As there is a difficulty of drug penetration into the brain due to the blood–brain barrier (BBB), many CNS drugs treatments fail in clinical trials. Hence, there is a need to develop effective CNS drugs following strategies for delivery to the brain by better selecting them as early as possible during the drug discovery process. The use of in vitro BBB models has proved useful to evaluate the impact of drugs/compounds toxicity, BBB permeation rates and molecular transport mechanisms within the brain cells in academic research and early-stage drug discovery. However, these studies that require biological material (animal brain or human cells) are time-consuming and involve costly amounts of materials and plastic wastes due to the format of the models. Hence, to adapt to the high yields needed in early-stage drug discoveries for compound screenings, a patented well-established human in vitro BBB model was miniaturized and automated into a 96-well format. This replicate met all the BBB model reliability criteria to get predictive results, allowing a significant reduction in biological materials, waste and a higher screening capacity for being extensively used during early-stage drug discovery studies.

Ellagitannins, Gallotannins and their Metabolites- The Contribution to the Anti-Inflammatory Effect of Food Products and Medicinal Plants

2019 ◽  
Vol 25 (37) ◽  
pp. 4946-4967 ◽  
Author(s):  
Anna K. Kiss ◽  
Jakub P. Piwowarski
Keyword(s):  
Immune Response ◽  
Gastrointestinal Tract ◽  
Gut Microbiota ◽  
Health Effects ◽  
Biological Effects ◽  
Food Products ◽  
Anti Inflammatory ◽  
The Impact

The popularity of food products and medicinal plant materials containing hydrolysable tannins (HT) is nowadays rapidly increasing. Among various health effects attributable to the products of plant origin rich in gallotannins and/or ellagitannins the most often underlined is the beneficial influence on diseases possessing inflammatory background. Results of clinical, interventional and animal in vivo studies clearly indicate the antiinflammatory potential of HT-containing products, as well as pure ellagitannins and gallotannins. In recent years a great emphasis has been put on the consideration of metabolism and bioavailability of natural products during examination of their biological effects. Conducted in vivo and in vitro studies of polyphenols metabolism put a new light on this issue and indicate the gut microbiota to play a crucial role in the health effects following their oral administration. The aim of the review is to summarize the knowledge about HT-containing products’ phytochemistry and their anti-inflammatory effects together with discussion of the data about observed biological activities with regards to the current concepts on the HTs’ bioavailability and metabolism. Orally administered HT-containing products due to the limited bioavailability of ellagitannins and gallotannins can influence immune response at the level of gastrointestinal tract as well as express modulating effects on the gut microbiota composition. However, due to the chemical changes being a result of their transit through gastrointestinal tract, comprising of hydrolysis and gut microbiota metabolism, the activity of produced metabolites has to be taken into consideration. Studies regarding biological effects of the HTs’ metabolites, in particular urolithins, indicate their strong and structure-dependent anti-inflammatory activities, being observed at the concentrations, which fit the range of their established bioavailability. The impact of HTs on inflammatory processes has been well established on various in vivo and in vitro models, while influence of microbiota metabolites on silencing the immune response gives a new perspective on understanding anti-inflammatory effects attributed to HT containing products, especially their postulated effectiveness in inflammatory bowel diseases (IBD) and cardiovascular diseases.

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