The production of genetically engineered pigs as xenotransplant donors aims to solve the severe shortage organs for transplantation in humans. first barrier successful xenotransplantation is hyperacute rejection (HAR). HAR a rapid and massive humoral immune response directed against pig carbohydrate Galα1,3-Gal epitope, which synthesized by α1,3-galactosyltransferase (α1,3-GT). antigen also contributes subsequent acute vascular events. Genetic modifications donor transgenic human complement...

Abstract Cloned pigs were produced from cultured skin fibroblasts derived a H‐transferase transgenic boar. One 90 day fetus and two healthy piglets resulted nuclear transfer by fusion of with enucleated oocytes. The cells used in these studies subjected to an extensive culture time, freezing thawing, clonal expansion single prior transfer. PCR FACS analysis determined that the cloned offspring contained expressed transgene. Microsatellite confirmed clones genetically identical cell...

Chromatin reorganization governs the regulation of gene expression during preimplantation development. However, landscape chromatin dynamics in this period has not been explored bovine. In study, we constructed a genome-wide map accessible bovine oocytes and early embryos using an improved assay for transposase-accessible with high-throughput sequencing (ATAC-seq) which revealed unique features embryo We found that accessibility is low 2-/4-cell embryos, followed by significant increase...

Supporting healthy pregnancy outcomes requires a comprehensive understanding of the molecular and cellular programs peri-implantation development, when most failure occurs. Here, we present single-cell transcriptomes bovine embryo development at day 12, 14, 16, 18 post-fertilization. We defined composition gene expression embryonic disc, hypoblast, trophoblast lineages in embryos, identified markers pathway signaling that represent distinct stages lineages; selected was validated embryos....

One-cell cattle embryos were prepared by in vitro oocyte maturation and fertilization (IVM/IVF) cultured with or without oviductal cells. Embryos evaluated after 7 days culture to determine the percentage developing from 1-cell stage morula blastocyst stage. The combination of glycine (2 mM) alanine (1 cells (experiment 1) improved embryo development over that control (29 vs. 13%; p < 0.05). An optimum response was obtained 10 mM 1 coculture (experiments 2 3). In experiment 4, effects (0...

Hyperacute rejection (HAR) is the first critical immunological hurdle that must be addressed in order to develop xenogeneic organs for human transplantation. In area of cell-based xenotransplant therapies, natural antibodies (XNA) and complement have also been considered barriers successful engraftment. Transgenic expression inhibitors donor cells has significantly prolonged survival xenografts. However, without eliminating antibody reactivity may provide insufficient protection clinical...

In vitro matured (IVM) bovine oocytes were examined to determine their potential viability in embryo cloning. Activation competence, as monitored by pronuclear formation, increased with oocyte age. Oocytes readily formed a pronucleus when challenged an electrical pulse 30 h after the onset of maturation. Developmental competence IVM tended increase age (P = 0.079). Selection on basis presence polar body 24 maturation and size follicle from which was derived improved development nuclear...

Summary. Bovine oocytes were bisected, stained with Hoechst 33342 and observed under a fluorescent microscope to identify nucleated enucleated demi-oocytes. Other bisected but not stained, or only half of each oocyte viewed microscope. The then used for nuclear transfer by fusing them embryonic blastomeres from 5–6 day bovine embryo. fusion rate proportion developing into compact morulae blastocysts was compared among different types Expt 1 examined the effect staining indicated no on either...

Abstract Micromanipulation and electrofusion were utilized for nuclear transfer in bovine embryos. Embryonic blastomeres from 5‐day (estrus = day 0), 6‐day, frozen‐thawed 5‐day, first‐generation embryos (embryos themselves a product of with the original donor being embryo) transferred into bisected oocytes by electrofusion. The percentage cells fusing recipient was compared between different types developing normally morula or blastocysts following 6 days culture sheep oviduct also recorded...

&lt;i&gt;Background:&lt;/i&gt; Stem cell characteristics such as self-renewal, differentiation and expression of CD34 CD44 stem markers have not been identified in porcine adipose tissue-derived adult (ADAS) cells. The objective this study was to develop a protocol for the isolation culture cells determine cell-like characteristics. &lt;i&gt;Methods:&lt;/i&gt; Primary cultures were established maintained. Cloning capacity determined using ring cloning procedure. clones differentiated stained...

Evidence indicates that failure of nuclear transfer (NT) embryos to develop normally can be attributed, at least partially, the use differentiated cells as donor karyoplast. Blastocyst production and development term cloned has been hypothesized differ between population doublings same cell line a consequence changes in levels DNA methyltransferase 1 (DNMT1) methylated during vitro culture. The objective this study was determine embryo production, developmental potential, gene expression...

Heat stress affects oocyte developmental competence and is a major cause of reduced fertility in heat stressed cattle. Negative effects on the have been observed at morphological, biochemical levels. However, mechanisms by which transcriptional epigenetic levels remain to be further elucidated. Here we aimed investigate effect quality, transcriptomic profiles DNA methylation oocytes collected through transition from spring summer under Louisiana conditions. Summer season resulted lower...

Abstract: Research in pig‐to‐primate xenotransplantation aims to solve the increasing shortage of organs for human allotransplantation and develop new cell‐ tissue‐based therapies. Progress towards its clinical application has been hampered by presence xenoreactive natural antibodies that bind foreign cell surface activate complement, causing humoral graft rejection. Genetic engineering donor cells animals express complement inhibitors such as hCD59 significantly prolonged survival....

Summary Embryo recovery rates from commercial donor mares mated at the clients' farms and research donors authors' laboratory were 27, 59 57 per cent, respectively. Transfer of embryos recovered on Days 6 to 8 post ovulation recipients that −1, −2 or −3 days asynchronous with respective gave pregnancy 54, 40 60 cent No pregnancies produced transfer 9 10 donors.