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Topic:Cell Proliferation
Cell proliferation in horses refers to the process by which cells divide and multiply, contributing to growth, development, and tissue repair. This biological process is fundamental to maintaining normal physiological functions and responding to injuries or diseases. In equine research, cell proliferation is studied to understand its role in various contexts such as wound healing, regenerative medicine, and cancer. Factors influencing cell proliferation in horses include genetic, environmental, and nutritional elements. This page assembles peer-reviewed research studies and scholarly articles that explore the mechanisms, regulation, and implications of cell proliferation in equine health and disease management.
In vitro heterogeneity of osteogenic cell populations at various equine skeletal sites. Bone cell cultures were evaluated to determine if osteogenic cell populations at different skeletal sites in the horse are heterogeneous. Osteogenic cells were isolated from cortical and cancellous bone in vitro by an explant culture method. Subcultured cells were induced to differentiate into bone-forming osteoblasts. The osteoblast phenotype was confirmed by immunohistochemical testing for osteocalcin and substantiated by positive staining of cells for alkaline phosphatase and the matrix materials collagen and glycosaminoglycans. Bone nodules were stained by the von Kossa method and counted....
Cell growth characteristics and differentiation frequency of adherent equine bone marrow-derived mesenchymal stromal cells: adipogenic and osteogenic capacity. To characterize equine bone marrow (BM)-derived mesenchymal stem cell (MSC) growth characteristics and frequency as well as their adipogenic and osteogenic differentiation potential. Methods: In vitro experimental study. Methods: Foals (n=3, age range, 17-51 days) and young horses (n=5, age range, 9 months to 5 years). Methods: Equine MSCs were harvested and isolated from sternal BM aspirates and grown up to passage 10 to determine cell-doubling (CD) characteristics. Limit dilution assays were performed on primary and passaged MSCs to determine the frequency of colony-forming units with a fibr...
Derivation and induction of the differentiation of animal ES cells as well as human pluripotent stem cells derived from fetal membrane. We succeeded in the derivation and maintenance of pluripotent embryonic stem (ES) cells from equine and bovine blastocysts. These cells expressed markers that are characteristics of mouse ES cells, namely, alkaline phosphatase, stage-specific embryonic antigen 1, STAT 3 and Oct 4. We confirmed the pluripotential ability of these cells, which were able to undergo somatic differentiation in vitro to neural progenitors and to endothelial or hematopoietic lineages. We were able to use bovine ES cells as a source of nuclei for nuclear transfer and we generated cloned cattle with a higher frequency ...
Horse embryonic stem cell lines from the proliferation of inner cell mass cells. Inner cell mass (ICM) cells were isolated immunosurgically from day 7-8 horse blastocysts and, after proliferation in vitro for 15-28 passages, three lines of cells were confirmed to be embryonic stem (ES) cells by their continued expression of alkaline phosphatase activity and their ability to bind antisera specific for the recognized stem cell markers, SSEA-1, TRA-1-60, TRA-1-81, and the key embryonic gene Oct-4. When maintained under feeder cell-free conditions in vitro, the three lines of cells differentiated into cells of ectodermal, endodermal, and mesodermal lineages. However, they did ...
Immunostimulatory effects of the anionic alkali mineral complex Barodon on equine lymphocytes. Previous studies have shown that the anionic alkali mineral complex BARODON has an immunoenhancing effect on pigs as an adjuvant and as a nonspecific immunostimulant. Likewise, the equine immune system has been defined with various monoclonal antibodies specific to equine leukocyte differentiation antigens to determine the possibility of enhancing equine resistance to respiratory diseases and promoting other immunostimulatory effects with the application of BARODON. Compared with the control group, after 3 weeks of treatment, BARODON-treated groups showed higher proportions of cells (P < 0.05)...
The equine periodontium as a continuously remodeling system: morphometrical analysis of cell proliferation. The hypsodont equine cheek tooth erupts continuously throughout life. Tooth eruption is inevitably associated with a remodeling of the periodontium. One major process of remodeling in the PDL is cell proliferation. The aim of this study was to detect cell proliferation at different sites of the equine PDL in order to examine the dynamics of the periodontal cell population. Methods: Specimens from nine warm-blood horses were taken-- containing the PDL interposed between the cementum and the alveolar bone--at three designated levels: subgingival, middle, and apical. Cell proliferation was detect...
Derivation, maintenance, and induction of the differentiation in vitro of equine embryonic stem cells. We describe here the isolation and maintenance of pluripotent embryonic stem (ES) cells from equine blastocysts that have been frozen and thawed. Equine ES cells appear to maintain a normal diploid karyotype in culture. These cells express markers that are characteristic of mouse ES cells, namely, alkaline phosphatase, stage-specific-embryonic antigen 1, STAT3, and Oct4. We also describe protocols for the induction of differentiation in vitro to neural precursor cells in the presence of basic fibroblast growth factor (bFGF), epidermal growth factor, and platelet-derived growth factor and to he...
Temporal relationship between proliferating and apoptotic hormone-producing and endothelial cells in the equine corpus luteum. The temporal relationship between endothelial cell death, vascular regression and the death of hormone-producing cells in the mare has not been established. To determine the dynamics of cell proliferation and death throughout the luteal phase, corpora lutea were studied at the early, mid- and late luteal phase, and after treatment with cloprostenol in the mid-luteal phase to induce premature luteolysis. Changes in cell proliferation and apoptosis were investigated utilising specific markers (phosphorylated histone-3 and activated caspase-3 respectively). Histone-3 positive cells were most abun...
Pathomorphological and immunohistochemical study of selected markers of tumour cell proliferation in equine sarcoids. The purpose of the study was a pathomorphological and immunohistochemical analysis of tumour cells and connective tissue in equine sarcoids. Investigations were performed using histopathological, ultrastructural, immunohistochemical (PCNA, p53, cytokeratin, vimentin) and histochemical (Ag-NORs) methods. The study was conducted on 50 sarcoids originating from 36 horses and classified as occult, verrucous, fibroblastic and a mixed type of sarcoid based on their clinical appearance. Most of the tumours were located on the girth (30%), neck (24%), head (12%), and legs (12%). The average age of the...
Signaling through the small G-protein Cdc42 is involved in insulin-like growth factor-I resistance in aging articular chondrocytes. During aging, chondrocytes become unresponsive to insulin-like growth factor-I (IGF-I). This study examined the role of Cdc42 (cell-division-cycle 42) in IGF-I signaling during aging. Experiments were performed using cartilage and chondrocytes isolated from horses ages 1 day-25 years. Northern analysis was used to examine expression of the small GTPases Cdc42, Rac, and RhoA. Western analysis was utilized to assess total Cdc42 (GTP + GDP-bound); active, GTP-Cdc42 was assessed using a pulldown assay with Western analysis. GTP-Cdc42 was also measured following IGF-I treatment. Gene expression for...
Equine peripheral blood-derived progenitors in comparison to bone marrow-derived mesenchymal stem cells. Fibroblast-like cells isolated from peripheral blood of human, canine, guinea pig, and rat have been demonstrated to possess the capacity to differentiate into several mesenchymal lineages. The aim of this work was to investigate the possibility of isolating pluripotent precursor cells from equine peripheral blood and compare them with equine bone marrow-derived mesenchymal stem cells. Human mesenchymal stem cells (MSCs) were used as a control for cell multipotency assessment. Venous blood (n = 33) and bone marrow (n = 5) were obtained from adult horses. Mononuclear cells were obtained by Fico...
Effect of fetal bovine serum and heat-inactivated fetal bovine serum on microbial cell wall-induced expression of procoagulant activity by equine and canine mononuclear cells in vitro. To determine the effect of fetal bovine serum (FBS) and heat-inactivated FBS (HI-FBS) on lipopolysaccharide (LPS)- and zymosan-induced procoagulant activity of equine and canine mononuclear cells. Methods: Mononuclear cells from 18 horses and 3 dogs. Methods: Cells were incubated with various concentrations of FBS, HI-FBS, LPS, zymosan, polymyxin B, and anti-LPS-binding protein monoclonal antibody or combinations of these constituents. A 1 stage recalcification assay was used to determine procoagulant activity. Results: Addition of FBS to media significantly increased procoagulant activity; eq...
Heaves, an asthma-like equine disease, involves airway smooth muscle remodeling. Increased airway smooth muscle mass is a prominent feature of asthmatic airway remodeling. Airway smooth muscle hyperplasia occurs in rodent models of experimental asthma, but the relevance of such finding to spontaneously occurring disease in large mammals is unknown. Objective: We examined horses with heaves, a naturally occurring equine asthma related to sensitization and exposure to moldy hay. We hypothesized that airway remodeling occurs in heaves and shares disease mechanisms with asthma. Methods: We quantified the airway smooth muscle mass and the numbers of proliferating and apoptotic ...
New staining methods for sperm evaluation estimated by microscopy and flow cytometry. New staining methods and automated instruments are now available to evaluate the sperm cell in vitro. Individual compartments of the sperm cell, such as the nucleus and the plasma and acrosomal membranes, may be investigated, as well as the cell function as shown by mitochondria activity and capacitation. Various probes are used and they can be analyzed by direct light or fluorescent microscopy or by flow cytometry. The automated instruments allow objective and accurate analysis and quantification as well as the ability to evaluate large population of cells in a shorter time, thus providing ac...
Blastocyst development in equine oocytes with low meiotic competence after suppression of meiosis with roscovitine prior to in vitro maturation. This study was conducted to evaluate the in vitro development of equine oocytes with compact cumuli that had been subjected to a period of meiotic suppression with roscovitine before in vitro maturation. In experiment 1, oocytes were recovered from slaughterhouse-derived ovaries and held in M199 + 10% fetal bovine serum containing 66 microM roscovitine with or without an overlay of mineral oil in 5% CO2 in air at 38.2 degrees C for 16-18 or 24 h. No oocytes treated with roscovitine in the absence of an oil overlay for 16-18 h were maturing, compared with 2-4% of oocytes in other treatments. In...
Inhibition of caspase-9 reduces chondrocyte apoptosis and proteoglycan loss following mechanical trauma. Chondrocyte death, a notable feature of osteoarthritis, may play a role in the initiation of cartilage degeneration. The present study was aimed at uncovering the nature and involvement of cell death in the initiation of cartilage degeneration induced by mechanical trauma. Methods: Articular cartilage discs obtained from healthy skeletally mature horses were subjected to a single-impact load (500 g from 50 mm) using a simple drop-tower device and cultured in vitro for 48 h. Chondrocyte death was examined using two independent methods: transmission electron microscopy and caspase-3 activity ass...
Anabolic effects of acellular bone marrow, platelet rich plasma, and serum on equine suspensory ligament fibroblasts in vitro. The purpose of this study was to investigate the response of suspensory ligament fibroblasts (SLF) to in vitro stimulation using acellular bone marrow (ABM), platelet rich plasma (in vitro PRP), and serum as potential treatment modalities for suspensory desmitis. Blood, bone marrow, and suspensory ligaments were collected from five horses. SLF were harvested, grown until confluent, and stimulated with various concentrations of ABM, PRP, equine serum, foetal bovine serum, and medium (control). The responses to the treatments were assessed using a combination of radio-labeling for total protein ...
Immune response to Sarcocystis neurona infection in naturally infected horses with equine protozoal myeloencephalitis. Equine protozoal myeloencephalitis (EPM) is one of the most common neurologic diseases of horses in the United States. The primary etiologic agent is Sarcocystis neurona. Currently, there is limited knowledge regarding the protective or pathophysiologic immune response to S. neurona infection or the subsequent development of EPM. The objectives of this study were to determine whether S. neurona infected horses with clinical signs of EPM had altered or suppressed immune responses compared to neurologically normal horses and if blood sample storage would influence these findings. Twenty clinical...
Effect of microcurrent electrical tissue stimulation on equine tenocytes in culture. To determine effects of microcurrent electrical tissue stimulation (METS) on equine tenocytes cultured from the superficial digital flexor tendon (SDFT). Methods: SDFTs were collected from 20 horses at slaughter. Methods: Tenocytes were isolated following outgrowth from explants and grown in 48-well plates. Four methods of delivering current to the tenocytes with a METS device were tested. Once the optimal method was selected, current consisting of 0 (negative control), 0.05, 0.1, 0.5, 1.0, or 1.5 mA was applied to cells (8 wells/current intensity) once daily for 8 minutes. Cells were treated ...
Serotonin-containing cells in the gastrointestinal tract of newborn foals and adult horses. Serotonin (5-hydroxytryptamine, 5-HT), a regulatory amine of mucosal enterochromaffin cells plays an important role in the control of gastrointestinal smooth muscle contraction and epithelial secretion. Serotonin has also been associated with gastric ulcers, diarrhoea and abdominal pain. In spite of the high incidence of these gastrointestinal disorders in newborn foals and adult horses, no data are available regarding 5-HT immunoreactive cells (i.c.) in the gastrointestinal tract (GIT) of foals, and for adult horses, data are incomplete and contradictory. In this study, the distribution and r...
Equine CCL11 induces eosinophil cytoskeletal reorganization and activation. To assess the biological effects of purified recombinant equine CCL11 on equine eosinophil function. Methods: Following stimulation of eosinophils from normal horses, the polymerised form of actin was measured by flow cytometry using fluorescently labelled phalloidin. Migration was determined in a 96 well plate chemotaxis assay using 8 microm pore membranes, and adherence of eosinophils to serum-coated plastic was assessed using a colorimetric assay for eosinophil peroxidase. Superoxide generation was measured by the reduction of cytochrome C in a colorimetric assay. Results: Equine CCL11 indu...
Progesterone receptors and proliferating cell nuclear antigen expression in equine luteal tissue. Steroid hormones act via specific receptors, and these play an important physiological role in the ovary. The objective of this study was to evaluate the cellular distribution of progesterone receptors and their staining intensity in different equine luteal structures during the breeding season, as well as their relationship to luteal cell composition, cell proliferation pattern and plasma progesterone (P4) concentration. There was an increase in proliferating cell nuclear antigen (PCNA) expression in large luteal cells from the corpus hemorrhagicum (CH) to mid-luteal phase, followed by a decr...
Somatic cell nuclear transfer in horses: effect of oocyte morphology, embryo reconstruction method and donor cell type. The objective of the present work was to investigate and clarify the factors affecting the efficiency of somatic cell nuclear transfer (NT) in the horse, including embryo reconstruction, in vitro culture to the blastocyst stage, embryo transfer, pregnancy monitoring and production of offspring. Matured oocytes, with zona pellucida or after zona removal, were fused to cumulus cells, granulosa cells, and fetal and adult fibroblasts, and fused couplets were cultured in vitro. Blastocyst development to Day 8 varied significantly among donor cells (from 1.3% to 16%, P < 0.05). In total, 137 nuclear...
The influence of 1-10 kD fraction from brains of the hibernating ground squirrel and the Yakut horse on proliferation and protein synthesizing system of Ehrlich ascitic carcinoma cells. The experimental data presented in the work testify to the cytostatic activity of 1-10 kD polypeptide fractions from brains of the hibernating ground squirrel and the Yakut horse towards Ehrlich ascitic carcinoma (EAC) cells. The experiments on the investigation of the inhibiting influence of 1-10 kD fractions from tissues of the hibernating and cold-adapted animals on protein-synthesizing system of EAC cells allow us to conclude that the cytostatic effect of the fractions is effected at the genetic level in the tumor cells.
Adding cholesterol to the stallion sperm plasma membrane improves cryosurvival. Cryopreservation induces partially irreversible damage to equine sperm membranes. Part of this damage occurs due to membrane alterations induced by the membrane changing from the fluid to the gel-state as the temperature is reduced lower than the membrane transition temperature. One way to prevent this damage is to increase the membrane fluidity at low temperatures by adding cholesterol to the membrane. Different concentrations of cholesterol-loaded-cyclodextrins (CLC) were added to stallion sperm to determine the CLC concentration that optimizes cryosurvival. Higher percentages of motile sper...
Microvascularization and angiogenic activity of equine corpora lutea throughout the estrous cycle. Corpus luteum growth and endocrine function are closely dependent on the formation of new capillaries. The objectives of this study were to evaluate (i) tissue growth and microvascular development in the equine cyclic luteal structures; (ii) in vitro angiogenic activity of luteal tissues in response to luteotrophic (LH, PGE(2)) and luteolytic (PGF(2alpha)) hormones and (iii) to relate data to luteal endocrinological function. Our results show that microvascular density was increased in the early and mid luteal phase, followed by a fall in the late luteal phase and a further decrease in the cor...
Effect of the in vitro maturation medium on equine oocytes: comparison of follicular fluid and oestrous mare serum. The present study evaluated the effect of supplementing the medium used to mature equine oocytes in vitro with oestrous mare serum (EMS) or horse follicular fluid (HFF). To this end, 144 ovaries were obtained from mares aged 16-21 months and transported to the laboratory in Dulbecco's phosphate buffered saline (D-PBS) at 30 degrees C. Oocytes were harvested from the ovaries by slicing, and then selected for in vitro maturation (IVM) according to the number of cumulus cell layers and the characteristics of the cytoplasm. The selected oocytes were washed three times in TCM199 medium plus HEPES (...
Effect of cryopreservation on the cellular integrity of equine embryos. Horse embryos are rarely cryopreserved in practice because expanded blastocysts tolerate freezing poorly, and the embryo begins expanding very soon after entering the uterine cavity. This study examined the effects of freezing on cytoskeleton integrity, and investigated whether cell damage could be reduced using trypsin to thin the blastocyst capsule or cytochalasin-B (cyto-B) to stabilise the cytoskeleton. Sixty-nine embryos were recovered 7 days after ovulation and equilibrated in 10% glycerol, with or without pretreatment with 0.2% trypsin or 7.5 microg/ml cyto-B. Forty-two of the embryos w...
