Development of Pyramidal Microwells for Enhanced Cell Spheroid Formation in a Cell-on-Chip Microfluidic System for Cardiac Differentiation of Mouse Embryonic Stem Cells.

Three-dimensional (3D) tissue culture models provide in vivo-like conditions for studying cell physiology. This study aimed to examine the efficiency of pyramidal microwell geometries in microfluidic devices on spheroid formation, cell growth, viability, and differentiation in mouse embryonic stem cells (mESCs). The static culture using the hanging drop (HD) method served as a control.

Novel electroporation microchip with field constriction enhances transfection efficiency and survival rates of feline embryos.

This study introduces a low-voltage electroporation microchip designed for transfection in cat embryos, featuring real-time impedance monitoring. The microchip uses a field constriction strategy, which localises the electric field to the membrane region in contact with the micro-orifice, enhancing electroporation efficiency while minimising damage. Embryos were positioned on the orifice, and a series of voltage pulses (10, 15, and 20 V) were applied.

Comparison of Fine-Needle Aspiration and Core Needle Biopsy for the Pre-Operative Diagnosis of Canine and Feline Mammary Gland Tumours.

Mammary gland tumours are common neoplasms that affect female dogs and cats. We compared the accuracy of pre-surgical fine-needle aspiration (FNA) and core needle biopsy (CNB) diagnosing feline (n = 64) and canine (n = 83) mammary gland tumours with excisional histopathology as the gold standard for the definitive diagnosis. We also explored the impact of CNB needle sizes (18G and 16G).

Heat Shock Related Protein Expression in Abdominal Testes of Asian Elephant ().

The abdominal testes of Asian elephants show normal spermatogenesis. Heat shock in cryptorchid testes elevates heat shock factor (HSF) expression, leading to germ cell apoptosis, while increased heat shock proteins (HSPs) levels provide protection. To investigate how heat shock affects elephant spermatogenic cells, focusing on heat shock-related molecules and the cell death mechanism, immunohistochemistry and TUNEL staining were employed to assess the immunoexpression of several heat shock-related molecules and the status of apoptosis in elephant fibroblasts (EF) induced by heat shock stimulus.

Modulation of MHC expression by interferon-gamma and its influence on PBMC-mediated cytotoxicity in canine mast cell tumour cells.

Immunotherapy is a promising alternative treatment for canine mast cell tumour (MCT). However, evasion of immune recognition by downregulating major histocompatibility complex (MHC) molecules might decline treatment efficiency. Enhancing MHC expression through interferon-gamma (IFN-γ) is crucial for effective immunotherapy.

Analysis of serum peptidome profiles of non-metastatic and metastatic feline mammary carcinoma using liquid chromatography-tandem mass spectrometry.

Background: Feline mammary carcinoma (FMC) is a common aggressive and highly metastatic cancer affecting female cats. Early detection is essential for preventing local and distant metastasis, thereby improving overall survival rates. While acquiring molecular data before surgery offers significant potential benefits, the current protein biomarkers for monitoring disease progression in non-metastatic FMC (NmFMC) and metastatic FMC (mFMC) are limited.

De novo reconstruction of a functional in vivo-like equine endometrium using collagen-based tissue engineering.

To better understand molecular aspects of equine endometrial function, there is a need for advanced in vitro culture systems that more closely imitate the intricate 3-dimensional (3D) in vivo endometrial structure than current techniques. However, development of a 3D in vitro model of this complex tissue is challenging. This study aimed to develop an in vitro 3D endometrial tissue (3D-ET) with an epithelial cell phenotype optimized by treatment with a Rho-associated protein kinase (ROCK) inhibitor.

Effects of freeze-drying on the quality and fertilising ability of goat sperm recovered from different parts of the epididymis.

Lyophilisation is an alternative method for sperm preservation. The aim of this study was to evaluate the effects of freeze-thawing (F/T) and freeze-drying (F/D) on the quality of epididymal goat sperm. Sperm from each region of the epididymis (caput, corpus and cauda) were collected and evaluated for the expression of phospholipase C zeta (PLC-ζ), protamine 1 (PRM1), transition protein 1 (TNP1) and 2 (TNP2).

Dietary Thw. improves reproductive performance of aged roosters.

The application of artificial insemination is particularly, owing to which breeder animals are considered an important resource in breeding farms. However, the reproductive performance of roosters typically declines with age, and the economic loss experienced by breeders is attributable to this shortened reproductive lifespan. Thw.

Dietary Thw. Improves Growth Performance in Broilers.

This study aimed to evaluate the effects of dietary Thw. (LST) powder supplementation on growth performance, blood metabolites, antioxidant status, intestinal morphology, and cecal microbiome in broiler chickens. A total of 400 1-day-old male Guangxi partridge broilers (initial body weight: 42.

Expression of oxytocin receptors and oxytocin assisted electroejaculation in the domestic cat (Felis catus).

Oxytocin is a peptide hormone that mainly functions to control the contractility of smooth muscles and sex-related steroidogenesis in male reproductive tracts. However, specific information concerning this hormone in controlling the reproductive organs of cats is limited. This study aimed to investigate the expression of oxytocin receptors (OTRs) and their signal mediator via prostacyclin synthase (PTGIS) in reproductive structures following oxytocin assisted electroejaculation.

Membrane-permeable trehalose improves the freezing ability and developmental competence of in-vitro matured feline oocytes.

Oocytes are highly sensitive to cryopreservation, which frequently results in an irreversible loss of developmental competence. We examined the effect of membrane-permeable trehalose on the freezing ability of feline oocytes matured in vitro. In Experiment 1, intracellular trehalose (trehalose hexaacetate; Tre-(OAc)) was synthesized from trehalose precursor and subjected to spectroscopic characterization.

Aberrant levels of DNA methylation and H3K9 acetylation in the testicular cells of crossbred cattle-yak showing infertility.

Although the interspecies hybridization of bovids, such as cattle-yak (Bos taurus × Bos grunniens), has heterosis benefits, the infertility of hybrid males affects the maintenance of dominant traits in subsequent generations. To achieve reproductive capacity, male germ cell development requires coordinated changes in gene expression, including DNA methylation and generalized histone modifications. Although gene expression-related mechanisms underlying hybrid male sterility have been investigated recently, information on the cell types and stage-specific controls remains limited.

Oocyte Cryopreservation in Domestic Animals and Humans: Principles, Techniques and Updated Outcomes.

Oocyte cryopreservation plays important roles in basic research and the application of models for genetic preservation and in clinical situations. This technology provides long-term storage of gametes for genetic banking and subsequent use with other assisted reproductive technologies. Until recently, oocytes have remained the most difficult cell type to freeze, as the oocytes per se are large with limited surface area to cytoplasm ratio.

Exogenous LIN28 Is Required for the Maintenance of Self-Renewal and Pluripotency in Presumptive Porcine-Induced Pluripotent Stem Cells.

Porcine species have been used in preclinical transplantation models for assessing the efficiency and safety of transplants before their application in human trials. Porcine-induced pluripotent stem cells (piPSCs) are traditionally established using four transcription factors (4TF): OCT4, SOX2, KLF4, and C-MYC. However, the inefficiencies in the reprogramming of piPSCs and the maintenance of their self-renewal and pluripotency remain challenges to be resolved.

Molecular signature and colony morphology affect in vitro pluripotency of porcine induced pluripotent stem cells.

Overall efficiency of cell reprogramming for porcine fibroblasts into induced pluripotent stem cells (iPSCs) is currently poor, and few cell lines have been established. This study examined gene expression during early phase of cellular reprogramming in the relationship to the iPSC colony morphology and in vitro pluripotent characteristics. Fibroblasts were reprogrammed with OCT4, SOX2, KLF4 and c-MYC.

On-farm lambing outcomes after transfer of vitrified and slow frozen embryos.

The aim of the present study was to determine the most suitable embryonic stage and embryo freezing technique for commercial implementation of frozen embryo trading by small-scale sheep producers. There was a 2 × 2 factorial design utilized for conducting the study consisting of two embryo stages (2-8 cells or morula/blastocyst) and two cryopreservation protocols (vitrification or slow-freezing). For the in vivo produced embryos, there were treatments of crossbred donor ewes to induce superovulation.

Optimal concentration of Rho-associated coiled-coil kinase (ROCK) inhibitor improved sperm membrane functionality and fertilizing ability of cryopreserved-thawed feline sperm.

Sperm cryopreservation induces irreversible loss of viability and fertilizing ability. This study aimed at examining the effects of Rho-associated, coiled-coil kinase (ROCK) inhibitor on quality of frozen-thawed feline sperm. Ejaculated semen from individual cats (n = 6) was examined for the expression of LIMK1 and LIMK2 mediated ROCK cascade.

Generation of porcine induced-pluripotent stem cells from Sertoli cells.

Induced pluripotent stem cells (iPSCs) are generated by reprogramming of somatic cells using four transcription factors: OCT4, SOX2, KLF-4, and c-MYC (OSKM). However, reprogramming efficiency of iPSCs is currently poor. In this study, we used the Sertoli line as a novel cell source for somatic cell reprogramming.

Treatment with chemical delipidation forskolin prior to cryopreservation improves the survival rates of swamp buffalo (Bubalus bubalis) and bovine (Bos indicus) in vitro produced embryos.

The cryopreservation of embryos is a technology developed for long-term genetic preservation. However, high sensitivity to low temperatures due to a large number of intracellular lipids within ruminant embryos compromises the success of this technique. The aim of this study was to examine the effects of using of lipolytic chemical agent forskolin, during in vitro producing of buffalo and bovine embryos on lipid contents, cryotolerance and subsequent developmental competence of these embryos.

Rabbit induced pluripotent stem cells retain capability of in vitro cardiac differentiation.

Stem cells are promising cell source for treatment of multiple diseases as well as myocardial infarction. Rabbit model has essentially used for cardiovascular diseases and regeneration but information on establishment of induced pluripotent stem cells (iPSCs) and differentiation potential is fairly limited. In addition, there is no report of cardiac differentiation from iPSCs in the rabbit model.

Generation of a pig induced pluripotent stem cell (piPSC) line from embryonic fibroblasts by incorporating LIN28 to the four transcriptional factor-mediated reprogramming: VSMUi001-D.

Pig induced pluripotent stem cell (piPSC) line was generated from embryonic fibroblast cells using retroviral transduction approaches carrying human transcriptional factors: OCT4, SOX2, KLF4, c-MYC and LIN28. The generated piPSC line, VSMUi001-D, was positive for alkaline phosphatase activity and expressed the pluripotency associated transcription factors including OCT4, SOX2, NANOG and surface markers SSEA-1, all iPSC hallmarks of authenticity. Furthermore, VSMUi001-D exhibited a normal karyotype and formed embryoid bodies in vitro and teratomas in vivo.

Effects of vitrification and a Rho-associated coiled-coil containing protein kinase 1 inhibitor on the meiotic and developmental competence of feline oocytes.

Oocyte cryopreservation is the technique of choice for the long-term storage of female gametes. However, it induces an irreversible loss of oocyte viability and function. We examined the effects of vitrification and a Rho-associated coiled-coil containing protein kinase 1 (ROCK1) inhibitor (ROCKi) on the meiotic and developmental competence of feline oocytes.

Establishment of a rabbit induced pluripotent stem cell (RbiPSC) line using lentiviral delivery of human pluripotency factors.

Rabbit Embryonic Fibroblast (RbEF) cells (from Hycole hybrid rabbit foetus) were reprogrammed by lentiviral delivery of a self-silencing hOKSM polycistronic vector. The pluripotency of the newly generated RbiPSC was verified by the expression of pluripotency-associated markers and by in vitro spontaneous differentiation towards the 3 germ layers. Furthermore, the spontaneous differentiation potential of the iPSC was also tested in vivo by teratoma assay.