Cell viability refers to the ability of cells to survive and function within their physiological environment. In horses, assessing cell viability is an important aspect of veterinary research, particularly in understanding the effects of various treatments, diseases, and environmental factors on equine cellular health. Techniques such as flow cytometry, trypan blue exclusion, and MTT assays are commonly used to evaluate cell viability in equine studies. These methods help determine the proportion of living cells in a sample, providing insights into cellular responses to different stimuli or conditions. This page compiles peer-reviewed research studies and scholarly articles that explore the methodologies, applications, and implications of cell viability assessments in equine research.
Johnson L.A new approach to quantification of Sertoli cells is described. The number of Sertoli cells per testis was calculated from the number of spermatids per testis, the number of spermatids per Sertoli cell apex, and the correction for the lifespan of spermatids enumerated per testis. To evaluate this method under different physiological conditions, testes from 28 adult (4-20-year) stallions obtained in the nonbreeding season (December-January) and from 28 adult stallions in the breeding season (June-July) were compared. Number of Sertoli cells per gram parenchyma was similar between seasons. Howev...
Garner DL, Pinkel D, Johnson LA, Pace MM.Spermatozoa from bulls, boars, dogs, horses, mice, and men were examined using a fluorogenic stain consisting of the membrane-permeant substrate carboxyfluorescin diacetate (CFDA) and the relatively membrane-impermeant nuclear stain propidium iodide (PI). Three distinct populations of spermatozoa were discernible in samples from each species upon microscopic examination. Individual spermatozoa, presumed to be viable because of their motility, retained products of the fluorescein chromophore throughout the cell. A second population of spermatozoa in which the nuclei stained red with PI retained...
Yoshihara E, Nakae T.In order to develop the cytotoxic liposome, the cytolytic effect of polycationic liposome was examined. Upon incubation of the stearylamine-containing liposome (stearylamine-liposome) with rabbit erythrocyte, a significant extent of hemolysis was observed. Hemolytic activity of the liposome depends on the amount of stearylamine in the liposome membrane. The plots of the initial rate of hemolysis versus the concentration of stearylamine-liposome showed a sigmoidal curve, suggesting that stearylamine-liposomes act cooperatively on the erythrocyte membrane. Hemolytic activity of stearylamine-lipo...
Kuypers FA, Easton EW, van den Hoven R, Wensing T, Roelofsen B, op den Kamp JA, van Deenen LL.The phospholipid composition and the distribution of phospholipids over the two leaflets of the membrane have been investigated for rabbit and horse erythrocyte membranes. Phosphatidylcholine (PC) comprises 39.4% and 41.3% of the total phospholipid complement of the rabbit and horse erythrocytes, respectively. In both membranes the distribution of this phospholipid is asymmetric: 70% of the PC is present in the outer layer of the rabbit membrane and 60% in that of the horse. The major species of this phospholipid class are the (1-palmitoyl-2-oleoyl)- and the (1-palmitoyl-2-linoleoyl)PC. The di...
Zink MC, Yager JA, Prescott JF, Wilkie BN.Bronchoalveolar lavage was performed 5 times, sequentially, on 3 healthy foals while each foal was 6 to 63 days of age. Phagocytosis and bactericidal assays were performed on recovered alveolar macrophages. Corynebacterium equi and alveolar macrophages at a ratio of 10:1 were incubated for 1 hour in medium containing 1% heat-inactivated rabbit anti-C equi serum. After incubation, greater than 90% of the alveolar macrophages contained at least 1 ingested bacterium and each alveolar macrophage contained 9.4 +/- 1.0 bacteria (mean +/- SE). After alveolar macrophages and C equi were incubated for ...
Richer CL, Romagnano A.Both dynamic G-banding and cell synchronization produced by bromodeoxyuridine (BrdU), were applied to equine chromosomes. BrdU incorporated during the first half of the S-phase is taken up into the R-bands that are early replicating. These bands, which have incorporated BrdU, cannot contract as usual and remain elongated; only the other regions of the chromosome, i.e., the G-bands, contract normally and are sharply defined. BrdU also can be used for cell synchronization. The addition of BrdU in a high concentration, 15 hours before harvest, and its removal 11 hours later, has two effects: init...
Farrar RG, Klei TR.Strongylus edentatus was successfully cultured in vitro to the fourth larval stage (L4). Some growth continued for periods of 40-50 days at which time reductions in viability were observed in some of the culture systems tested. Various combinations of media, sera, buffers and organ explant cultures were tested. All cultures were incubated at 37 C in an atmosphere of 95% air and 5% CO2. Larvae underwent growth and differentiation to the L4 in all medium-serum combinations with and without organ explant cultures. Development and growth did occur but viability was reduced to insignificant levels ...
Broström H, Obel N, Perlmann P.Human lymphocytes displayed a frequent natural cytotoxicity (NK) in vitro against normal equine dermal fibroblasts (ED) and against equine tumour cells of a virus-containing cell line (Mc-1). Similarly, human normal sera contained antibodies that induced antibody-dependent cellular cytotoxicity (ADCC) by normal human lymphocytes against the same target cells. Both NK and ADCC varied for different donors. For individual donors, however, cytotoxicity against the two target cells was significantly correlated both in NK and ADCC. For ED there was also a significant correlation between ADCC and NK ...
Jørgensen RJ, Andersen S.Between 10 and 25% of the Dictyocaulus arnfieldi larvae excreted in faeces from a naturally infected donkey were harvested as infective stages from faecal cultures by means of Pilobolus fungi. The faeces were collected between 24 and 56 hours after drenching the donor animal with Pilobolus spores and kept at 16 +/- 2 degrees C. Most larvae were collected between the 5th and the 8th day of culturing during which period fructification and sporangium discharge also peaked. The sporangia and the adhering larvae were collected in Petri dishes inserted between the faecal mass and a light source. All...
Hegre OD, Marshall S, Hickey GE.Plasma amine oxidase activities (benzylamine oxidase and spermine oxidase) were determined in the sera of a number of species of various ages. Benzylamine oxidase (BZO) activity, measured spectrophotometrically, was present in bovine, equine, and ovine species examined. Generally its activity in serum increased with the age of the animal. Spermine oxidase activity (SPO) was estimated by a bioassay of in vitro toxicity and did not necessarily correlate with BZO. Cytotoxicity in the presence of spermidine was found only in the sera of the ruminant species examined. Serum activity tended to rise ...
Dyer RM, Liggitt HD, Leid RW.A device was constructed from an equine nasogastric tube, polyethylene tubing, and a 3-way stopcock and used to lavage the lungs of anesthetized ponies. The technique was safe and atraumatic in that 6.4 to 19.7 X 10(7) purified alveolar macrophages were removed from the lungs without harm to the ponies or contamination of the samples with blood. Studies of these highly purified cell suspensions revealed a mean viability of 85% as assessed by eosin dye exclusion with a mean recovery (+/- SD) of 12.5 +/- 4.8 X 10(7) pulmonary alveolar macrophages/pony.
Valeri CR, Valeri DA, Gray A, Contreras TJ, Lindberg JR.When equine RBC were frozen with 20% (w/v) glycerol and stored at -150 C for as long as 5 years, there were no adverse effects on freeze-thaw or freeze-thaw-wash recovery or oxygen transport function. The manner in which the glycerol was added to, and removed from, the equine RBC was shown to be an important consideration in ensuring optimal freeze-thaw-wash recovery values.
Kirkland KD, Fales WH, Blanchard TL, Youngquist RS, Hurtgen JP.Five isolants of Pseudomonas aeruginosa collected from clinical cases of equine genital infection and one standard strain of P. aeruginosa were exposed to various concentrations of ethylene-diaminetetraacetic acid (EDTA) and tris (hydroxymethyl) aminomethane (tris buffer pH 8) and EDTA-tris lysozyme. Colony forming units of the isolants and minimal inhibitory concentrations for 11 antimicrobial agents were determined with each isolant before and after exposure to the EDTA solutions. Decreased cellular viability was found with all six isolants after exposure to the EDTA-tris solutions. Reversal...
Peters GJ, De Abreu RA, Oosterhof A, Veerkamp JH.Concentrations of purine and pyrimidine ribonucleotides were measured with HPLC in lymphocytes of man, horse, pig and sheep and in rat thymocytes. The ATP concentration was highest in lymphocytes of all species and about 850 pmol/10(6) cells in human and equine lymphocytes, higher in porcine and lower in ovine lymphocytes and rat thymocytes. The GTP concentration was comparable in human, equine and porcine lymphocytes, but lower in ovine lymphocytes. ATP concentration was also measured in lymphocytes of man, horse and pig with a luciferin-luciferase assay. During culturing with or without phyt...
Bruyninckx WJ, Blancquaert AM.Horse mononuclear cells were separated from whole blood using neutral density gradient centrifugation on Isopaque-Ficoll. The resulting cell suspension was comparable in composition with similarly prepared human and bovine mononuclear cell preparations. The relative concentration of monocytes was increased by the use of a gradient with density lower than that originally proposed by Böyum (Böyum, A. 1968. Scand. J. Clin. Lab. Investig. 21 supple. 97:77-89). Contamination by neutrophils was limited either by using a gradient medium of lower density or by replacing Isopaque-Ficoll by Percoll-0....
Jacobsen K, Gintz T, Reed SM, Newbry J, Bayly WM, Perryman LE, Leid RW.Equine neutrophils (PMN) were isolated to greater than 99% purity by isopyknic sedimentation on coated colloidal silica particles. A cell recovery of 84.7 +/- 4.0%, with a viability of greater than 99%, was observed with this method. The isolated PMN were compared with mixed population of equine peripheral leukocytes with respect to functional integrity by chemiluminescence and bactericidal assays. There was no significant difference (P less than 0.01) observed in either assay between the isolated equine PMN and the mixed-cell populations. The methods used in both the isolation as well as the ...
Camp CJ, Leid RW.A method for the isolation of equine neutrophils was developed using metrizamide cushions. A purity of greater than 95% was routinely obtained with greater than 90% viability. These cells were radiolabeled and tested for their chemotactic response in Boyden chambers to zymosan-activated equine serum, the partially purified equine complement component C5a, and formyl-L-methionyl-L-leucyl-L-phenylalanine. The time and ionic requirements for chemotaxis of radiolabeled equine neutrophils were investigated and maximal movement was observed at 2 hours' incubation and 1.0 mM Ca and 0.5 mM Mg. Dinitro...
Doyle A, Owen LN.Sixteen cytotoxic drugs used in cancer chemotherapy in man were studied for cytopathic effect on equine fibrosarcoma, melanoma and normal equine lung cells in vitro. Three drugs, vincristine, melphalan and methotrexate, produced cytopathic effect
Harry TO, McCollum WH.The Bucyrus strain of equine arteritis virus, previously modified to avirulence and vaccinal virus by 131 serial passages in primary cell cultures of horse kidney followed by 111 passages in primary cell cultures of rabbit kidney, was further passaged in cultures of the E. Derm (NBL-6) cell line, a continuous diploid cell line. Pools of the 16th and 25th passages of the virus in this last equine dermal cell line were lyophilized and stored in lots at 37 C, 23 to 28 C, 4C, and -20 C. The viability of the vaccinal virus deteriorated rapidly during storage at 37 C and at 23 to 28 C, but was relat...
Dutta SK, Bumgardner MK, Scott JC, Myrup AC.Various methods of separation and identification of major equine leukocyte populations and subpopulations were used. The purity of T and B lymphocytes separated in Sephadex anti-equine F(ab')2 columns was 87% to 99% and 83% of 97%, respectively. The purity of T lymphocytes separated in nylon-wool columns was 89% to 98%. Preparations of B lymphocytes separated in glass-bead columns were 68% to 79% pure. The presence (or absence) of surface immunoglobulin by immunofluorescence was the most consistent and reliable method for the identification of B or T lymphocytes, respectively. However, the ery...
Carakostas MC, Moore WE, Smith JE.A procedure with chromium-51 (51Cr) as the cell label that maintains high-cell viability for studying granulocyte kinetics in horses is described. The procedure combines and modifies several methods for isolating leukocytes and granulocytes for use in the horse when a large volume of labeled cells is required. Also described is an improved technique for measuring granulocyte specific activity in large serial blood samples, using a Ficoll-sedimentation method. The procedure should be useful for determining granulocyte kinetics in the horse, the only major domestic species for which such data ar...
Glick DM, Mockel J.The disposition of 45Ca was studied in equine parathyroid gland slices. This preparation was shown to be viable by its ability to carry out protein synthesis, by the responsiveness of its respiration to effectors of oxidative phosphorylation, and by its calcium-inhibitable PTH secretion. 45Ca accumulates in the slices as the extracellular calcium is raised through the range in which physiological serum calcium concentration lies. The tissue slices behave as if they have no calcium pump: (a) depriving the cells of energy by treatment with antimycin A does not admit more calcium to the tissue, (...
Wilsman NJ, Farnum CE, Reed-Aksamit DK.The incidence and structure of equine and murine chondrocytic cilia were studied using serial sections and transmission electron microscopy. Overall, 96% of all equine chondrocytes and 100% of all murine chondrocytes had one cilium. The structure of these cilia included rootlets, basal feet, alar sheets, and an axoneme of nine peripheral doublets which progressively bent and terminated as they coursed towards the tip of the ciliary shaft. Together with the previous studies on neonatal and adult canine chondrocytic cilia, we conclude that the structure and incidence of chondrocytic cilia does n...
Ferrante A, Thong YH.Results presented show that highly purified populations of mononuclear (MN) and polymorphonuclear (PMN) leucocytes can be obtained from horse blood by a procedure similar to that previously described for the separation of these leucocytes from human blood. This involved centrifugation of horse blood on a Ficoll-Hypaque medium with a density of 1.095 g/ml. The procedure required approximately 1 h for completion and resulted in the simultaneous preparation of MN (greater than 98% purity) and PMN (greater than 96% purity) leucocytes. Cell viability exceeded 95% and cells retained immunological fu...
Verger C.We have shown previously that hemoglobin greatly stimulates chick embryo cell proliferation in Eagle's minimal essential medium supplemented with horse serum. In the present study we compared the effects of horse serum plus 10 micrometers hemoglobin to those of fetal bovine serum on subcultures of chick embryo cells serially propagated at high cell densities. The cells became elongated in the presence of fetal bovine serum and their rate of proliferation progressively decreased, whereas they became polygonal in the presence of horse serum plus hemoglobin and proliferated well in successive cel...
Kenney RM, Condon W, Ganjam VK, Channing C.The histological features and hormonal content of follicular fluid of antral follicles during oestrus were correlated. As a result it was possible to characterize several categories of viable and atretic follicles. A seemingly important stage in maturation appeared to be at 3 cm in diameter since follicular oestrogens and androgens underwent a 3-fold increase in concentration at that size. Evidence was obtained to suggest that oestrogens are anti-atretogenic. However, a drop in oestrogens was not the cause of atresia since degeneration commenced when levels were high. Contrary to the concept t...
Jörg A, Portmann P, Fellay G, Dreyer JL, Meyer J.An improved and short method is described for the isolation of intact eosinophilic leukocytes from horse blood with high yield (1--1.5 g/20 l). Viability and purity of the preparations were verified by light and electron microscopy and by the trypan blue exclusion test. Isolated eosinophils were 98--100% pure, intact and viable, and they could be shown to phagocytise immune-complexes.
Targowski SP.The present study describes a two step technique for the separation of mononuclear leukocytes or mononuclear and polymorphonuclear leukocytes from whole equine blood. First, the leukocyte rich plasma was obtained by sedimentation of erythrocytes in the undiluted blood. Subsequently, separation of the different populations of white blood cells was performed by centrifugation with different gradients overlaid with the leukocyte rich plasma. The optimal separation of the mononuclear cells was obtained by the centrifugation of the leukocyte rich plasma overlaying the gradient containing 24 parts o...
Broström H, Obel N, Perlmann P.Human lymphocytes displayed a frequent natural cytotoxicity (NK) in vitro against normal equine dermal fibroblasts (ED) and against equine tumour cells of a virus-containing cell line (Mc-1). Similarly, human normal sera contained antibodies that induced antibody-dependent cellular cytotoxicity (ADCC) by normal human lymphocytes against the same target cells. Both NK and ADCC varied for different donors. For individual donors, however, cytotoxicity against the two target cells was significantly correlated both in NK and ADCC. For ED there was also a significant correlation between ADCC and NK ...
Wu H, Han D, Jiang Z, Zhao D, Liu M, Xu X, Liu X, Yang L, Ji X, Wang M, Zhang S.B cell activating factor (BAFF) and its receptors were regarded as elements of the immune system, regulating the fate of B cell. In recent years, these molecules were identified in a number of normal and pathological tissues, expanding their potential functions beyond the immune system. In this study, on the basis of molecular clone and prokaryotic expression of equine BAFF, we reported that equine adipose-derived stem cell (ASC) expressed BAFF and its receptors, which exhibited the increased expression during ASC adipogenic differentiation in vitro. Moreover, with the addition of recombinant ...
Haysom LZ, Lee-Fowler TM, Spangler EA.Storage and temperature significantly impact bronchoalveolar lavage fluid (BALF) analysis, and shipment of samples to diagnostic laboratories is often necessary. Alternative sample preparation methods could limit storage and temperature effects. Objective: This study aimed to determine if airway wash samples that were fixed in formalin after being embedded in Histogel or Gelfoam gave comparable results to fresh cytocentrifuged or sediment smear preparations for the evaluation of cell morphology. Methods: Eleven bronchoalveolar lavage and 3 transtracheal wash fluids were available, including 8 ...
Glick DM, Mockel J.The disposition of 45Ca was studied in equine parathyroid gland slices. This preparation was shown to be viable by its ability to carry out protein synthesis, by the responsiveness of its respiration to effectors of oxidative phosphorylation, and by its calcium-inhibitable PTH secretion. 45Ca accumulates in the slices as the extracellular calcium is raised through the range in which physiological serum calcium concentration lies. The tissue slices behave as if they have no calcium pump: (a) depriving the cells of energy by treatment with antimycin A does not admit more calcium to the tissue, (...
Reedy SE, Powell DM, Williams NM, Dodson MV, Fitzgerald BP.This paper describes attempts to initiate equine adipocyte cultures from necropsy cases with varying intervals from time of death to isolation and culture. Equine adipocytes were isolated from 21 necropsy cases, regardless of the interval from time after death to establishment in primary ceiling cultures. However, while all cultures produced adipocytes, only 2 attempts to produce long-term equine adipocyte cultures from the subcutaneous rump fat depots were successful and not contaminated. Findings from these experiments indicate that it is possible to collect and culture equine adipocytes fro...
Schuetz AW, Whittingham DG, Legg RF.Granulosa cells at different stages of differentiation were collected from ovarian follicles and oviducts during the periovulatory period, and their nuclear DNA content was monitored by flow cytometry to establish their cell cycle characteristics (G0 + G1, S, G2 + M). The proportion of cells in the three phases of the cell cycle varied in characteristics patterns depending upon the time they were collected, before or following ovulation. Granulosa (cumulus) cells recovered from ovulated oocytes were mitotically inactive as shown by the large proportion of cells with a 2C amount of DNA and the ...
Lopez BS, Hurley DJ, Giancola S, Giguère S, Felippe MJB, Hart KA.The impact of culture conditions on equine monocyte-derived dendritic cells (MoDC) generation has not been fully characterized. We hypothesized that 1) MoDC could be cultured in a commercially available serum-free medium (AIM-V); and 2) that differential culture conditions would influence MoDC viability, yield and phenotype. MoDC generated from adult horses were cultured under variable conditions in a series of experiments. Viability was assessed using trypan blue and propidium iodide staining. Yield was determined by manual hemocytometer counting. Phenotype was assessed by flow cytometric ana...
Turn JT, Mayer J, Nagata K, Banovic F, Meichner K, Hurley DJ, Koslowski E, Gogal RM.Apitherapy is a form of alternative medicine that utilizes products from the western honeybee (Apis mellifera), including honey, propolis, and honeybee venom, to improve the health status of human patients by altering host immunity. An added benefit of these products is that they are nutraceuticals and relatively inexpensive to aquire. Currently, little is known about the use of honeybee products in veterinary species, as well as their impact on host immunity. In the present in vitro study, honey, propolis, and honeybee venom were co-cultured with enriched canine, equine, and chicken periphera...
Aguila L, Cabrera P, Arias ME, Silva M, Felmer R.Intracytoplasmic sperm injection (ICSI) in horses is currently employed for clinical and commercial uses, but the protocol could be optimized to improve its efficiency. We have hypothesized that destabilization of plasma and acrosomal membranes prior to injection would positively impact the developmental potential of equine zygotes generated by ICSI. This study evaluated effects of the sperm treatment with lysolecithin on plasma and acrosomal membranes and on oocyte activation ability, initially following heterologous ICSI on bovine oocytes and subsequently employing equine oocytes. The effect...
Giraldo A, Koch TG, Madan P, Lepage S, Monteith G, Alizadeh AH, Tran A, Mortagy N, Koenig JB.There is a knowledge gap regarding the effect of extracorporeal shockwave treatment (ESWT) on the stress response and immunomodulatory and anti-inflammatory properties of equine umbilical cord blood mesenchymal stromal cells (CB-MSCs). The objective of this study was to investigate the presence of cellular oxidative stress, inflammatory response, and production of growth factors in CB-MSCs after treatment with ESWT. We hypothesized that CB-MSCs treated with ESWT will experience higher levels of cellular stress and increased production of anti-inflammatory cytokines and growth factors compared ...
Jung H, Yoon M.Spermatogonial stem cells originate from gonocytes and undergo self-renewal and differentiation to generate mature spermatozoa via spermatogenesis in the seminiferous tubules of the testis in male mammals. Owing to the unique capacity of these cells, the spermatogonial stem cell transplantation technique, which enables the restoration of male fertility by transfer of germlines between donor and recipient males, has been developed. Thus, spermatogonial stem cell transplantation can be used as an important next-generation reproductive and breeding tool in livestock production. However, in large ...
Lessiak U, Melchert M, Walter I, Kummer S, Nell B, Tschulenk W, Pratscher B.Angiogenesis plays a crucial role in various physiological and pathological conditions. However, research in equine angiogenesis is relative limited, necessitating the development of suitable models. To effectively analyze angiogenesis in-vitro, it is essential to target the specific cells responsible for this process, namely endothelial cells. Human umbilical vein endothelial cells (HUVECs) are one of the most used models for studying angiogenesis in humans. Serving as an equivalent to HUVECs, we present a comprehensive isolation protocol for equine umbilical vein endothelial cells (EqUVECs...
Kader T, Lin JR, Hug C, Coy S, Chen YA, de Bruijn I, Shih N, Jung E, Pelletier RJ, Leon ML, Mingo G, Omran DK, Lee JS, Yapp C, Satravada BA, Kundra R....High-Grade Serous Ovarian Cancer (HGSOC) originates from fallopian tube (FT) precursors. However, the molecular changes that occur as precancerous lesions progress to HGSOC are not well understood. To address this, we integrated high-plex imaging and spatial transcriptomics to analyze human tissue samples at different stages of HGSOC development, including p53 signatures, serous tubal intraepithelial carcinomas (STIC), and invasive HGSOC. Our findings reveal immune modulating mechanisms within precursor epithelium, characterized by chromosomal instability, persistent interferon (IFN) signaling...
Luan Y, Zhong L, Li C, Yue X, Ye M, Wang J, Zhu Y, Wang Q.Polydactyly was recorded before 100 BCE and attracted widespread interest because of its relationship to limb health and ancestral traits in horses. However, the underlying reasons for the development of polydactyly remain unclear. To search for polydactyly-related genes, we utilize a paternal half-sib family and screen for variants that match the mode of inheritance. Through this screening process, 77 variants in 65 genes are filtered. A missense variant (EqCab3.0 chr4: <107353368> A > G) (rs1138485164) in the 3rd exon of LMBR1 is identified as a source of amino acid sequence variatio...
Bianchera A, Borghetti P, Ravanetti F, Bertocchi L, De Angelis E, Bettini R.Hyaluronic acid (HA) is a key component of synovial fluid as it plays a crucial role in joint physiology. Its biological activity is influenced by molecular weight, local concentration, and persistence in joints. High-molecular-weight HA has a consolidated history of clinical use, whereas little is known about the metabolic effect of low-molecular-weight hyaluronate on cartilage differentiation. This study explores the potential of HA-based nanoparticles (NPs) on chondrocytes differentiation in vitro. Starting from 25 kDa and 250 kDa sodium hyaluronate solutions, two types of NPs were prepared...
Ma Z, Li Y, Zhao Z, Song Q, Wang Q, Lu S, Wang J.In this study, we purified and identified antioxidant peptides from equine plasma protein hydrolysates and assessed their protective effects against HO-induced oxidative stress in Caco-2 cells. Four antioxidant peptides were identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS) in equine plasma protein hydrolysate, namely: GTMVGC (567.69 Da), FGMTST (662.88 Da), VGYHSHF (847.01 Da) and ALSPFFKE (939.18 Da). Among them, ALSPFFKE showed the strongest antidigestive properties after modelled digestion studies. Moreover, ALSPFFKE enhanced intracellular superoxide dismutase (SO...
Frontiers in geneticsFebruary 27, 2025
Volume 16 1534461 doi: 10.3389/fgene.2025.1534461
Barber AM, Kingsley NB, Peng S, Giulotto E, Bellone RR, Finno CJ, Kalbfleisch T, Petersen JL.The Functional Annotation of Animal Genomes (FAANG) consortium aims to annotate animal genomes across species, and work in the horse has substantially contributed to that goal. As part of this initiative, chromatin immunoprecipitation with sequencing (ChIP-seq) was performed to identify histone modifications corresponding to enhancers (H3K4me1), promoters (H3K4me3), activators (H3K27ac), and repressors (H3K27me3) in eight tissues from two Thoroughbred stallions: adipose, parietal cortex, heart, lamina, liver, lung, skeletal muscle, and testis. The average genome coverage of peaks identified by...
Castanheira CIGD, Anderson JR, Clarke EJ, Hackl M, James V, Clegg PD, Peffers MJ.This study describes a novel technique to analyze the extracellular vesicle (EV)-derived microRNA (miRNA) crosstalk between equine chondrocytes and synoviocytes. Donor cells (chondrocytes, n = 8; synoviocytes, n = 9) were labelled with 5-ethynyl uridine (5-EU); EVs were isolated from culture media and incubated with recipient cells (chondrocytes [n = 5] were incubated with synoviocyte-derived EVs, and synoviocytes [n = 4] were incubated with chondrocyte-derived EVs). Total RNA was extracted from recipient cells; the 5-EU-labelled RNA was recovered and sequenced. Differential expression analysi...
Baatar T, Song D, Weng Y, Wang G, Jin L, Guo R, Li B, Dugarjaviin M.The epigenetic regulation of gene expression through the covalent modification of histones is crucial for developing germline cells. To study the regulatory role of alternative splicing (AS) of euchromatic histone lysine methyltransferase 2 (EHMT2/G9A) in spermatogenesis in Mongolian horses, this study first examines the localization of the EHMT2 gene in testicular support cells and then predicts the higher-order structures of sequences with and without AS. Two types of lentiviral vectors for overexpression were subsequently constructed for the EHMT2 gene, one with AS and one without, to infec...
Cardinaux EM, Oltmanns H, Beineke A, Meißner J, Geburek F.Flexural limb deformities are a widespread condition in foals. Oxytetracycline is a common conservative treatment option with relaxing effects on the muscle-tendon-unit, potentially mediated through a matrix-metalloproteinase (MMP)-inhibitor mechanism. Its high therapeutic dose for this indication, potential negative side effects, and the guidelines for prudent use of antimicrobials make investigating alternatives desirable. In this study, the influence of substances with potentially similar mechanisms of action, however without antimicrobial properties, on viability and proliferation of juven...
Bosman LM, Ambele MA, Pepper MS.Regenerative medicine is a relatively new branch of therapeutics in equine medicine, which aims to restore and reconstitute tissue function and structure via cellular and/or noncellular approaches. Biological constituents such as mesenchymal stromal/stem cells (MSCs) are potent therapeutics, which can aid in damaged tissue regeneration due to their differentiation capacity into many different cell types such as adipose tissue, bone, and cartilage. MSCs can be successfully and conveniently isolated from equine subcutaneous adipose tissue (adipose-derived stromal cells, ASCs). In horses, there a...
Parsley AL, Hollingshead NA, Gruber EJ.To minimise storage-related artefacts, EDTA-anticoagulated whole blood should be evaluated promptly or refrigerated, a challenge in ambulatory practice. The effects of suboptimal blood storage on equine leukocyte parameters are not well-documented. Objective: Document changes in leukocyte number, automated and manual differential counts, and morphology in equine blood stored at four temperatures for up to 5 days. Methods: In vitro experimental. Methods: EDTA-anticoagulated whole blood from 13 horses was stored in a refrigerator (4°C), cooler with icepacks (3°C-19°C), room temperature (22...
Marine L, Wilfried P, Joëlle P, Justine J, Axel D, Vinciane T, Benoît V, Laurent G, Luc G, Jean-François K, Nadine A.In situ injection of mesenchymal stem cells appears as a promising treatment of tendinopathies. Tendon-derived mesenchymal stem cells (TDSCs) are widely studied and show a lot of interesting characteristics for clinical use. The aim of this study is to confirm the tenogenic potential of cryopreserved TDSCs and to confirm their ability to produce type I and/or type III collagens fibers in culture. Tendon-derived mesenchymal stem cells are harvested from the tendon no later than 72 h post-mortem. Their tenogenic potential has been assessed by quantitative reverse transcriptase polymerase chain ...
Contentin R, Jehl C, Commenchail K, Legendre F, Galéra P, Cassé F, Demoor M.Osteoarthritis (OA) affects millions of people globally, causing irreversible cartilage damage, chronic inflammation, and progressive joint dysfunction. Similarly, horses can develop OA spontaneously or due to their athletic careers, influenced by mechanical and biochemical factors. Current treatments primarily focus on symptom relief without promoting cartilage regeneration. In line with the 3Rs principles (refine, reduce, replace), the development of OA models is essential for advancing new therapeutic approaches against OA. In response to this need, the present study aimed to develop an m...
Hernández-Avilés C.Sperm quality analysis using computer-assisted sperm analysis (CASA) systems and fluorescence-based techniques has become common in the animal reproduction industry, particularly for large animals (i.e., bovine, porcine, equine). In this chapter, the methods commonly utilized in the author's laboratory to examine sperm motion characteristics via CASA and plasma membrane intactness by flow cytometry will be described. These include methods to properly dilute fresh (stallions, bulls, boars), cool-stored (stallions, boars), or frozen/thawed (stallions, bulls, boars) sperm for assessment of sperm ...
Semik-Gurgul E, Ocłoń E, Zubel-Łojek J, Pędziwiatr R, Pawlina-Tyszko K.Sarcoids are locally invasive skin tumors in equids, associated with bovine papillomavirus. Objective: Address potential applications of cannabidiol (CBD) in veterinary medicine. We evaluated the response of equine sarcoid cells to CBD in vitro, focusing on viability, invasiveness, and matrix remodeling. Methods: Three primary sarcoid cell lines. Methods: Cells were treated with CBD (20, 6.75, 2.25, 0.75 μM) and incubated for 6, 24, 48, 72 hours. Cell viability, cytotoxicity, and apoptosis were assessed using the ApoTox-Glo Assay. Based on these results, further analyses were performed for ...
Normélia P Morais A, Gastal GDA, Aguiar FLN, Hyde KA, Gastal MO, Lima LF, Ñaupas LVS, Flávia B Silva A, Becerra VAB, Gheno L, Curcio BR....Understanding how to effectively isolate a large number of healthy follicles is essential for increasing success rates in ARTs, such as in vitro culture and cryopreservation. Late secondary (preantral) and early tertiary (antral) follicles isolated from the intermediary portion of equine ovarian tissue sliced at two different thicknesses (0.25 mm vs. 0.50 mm) were characterized according to regional distribution (dorsal, medial, and ventral), morphofunctional features, and epigenetic profiles. After isolation via microdissection, follicles were evaluated for membrane integrity (trypan blue), v...
Kushida C, Tamura N, Kasashima Y, Sato K, Arai K.In equine regenerative medicine using bone marrow-derived mesenchymal stem/stromal cells (BM-MSC), the importance of the quality management of BM-MSC has been widely recognized. However, there is little information concerning the relationship between cellular senescence and the stemness in equine BM-MSC. In this study, we showed that stemness markers (NANOG, OCT4, SOX2 and telomerase reverse transcriptase) and colony forming unit-fibroblast apparently decreased accompanied with incidence of senescence-associated β-galactosidase-positive cells by repeated passage. Additionally, we suggested th...
Spacek SG, Hernández-Avilés C, Ramírez-Agámez L, Pearson M, Love CC.Cryopreservation of stallion epididymal sperm has become a common clinical procedure after routine castration, euthanasia, or acute death. Unique features of epididymal sperm compared to ejaculated sperm include the requirement to remove sperm from the cauda epididymis, lack of exposure to seminal plasma, and potential recovery of large sperm numbers. In this study, the effect of the flushing technique (Extender (INRA) vs. AIR), freezing extender (LE, CMLE, MFR5, CFR5, or BOTU), and the concentration at which sperm are cryopreserved (200, 400, or 800 × 10 sperm/mL) on post-thaw epididymal s...
