Recent Articles

Below are the titles and abstracts from the most recent articles published in In Vitro Cellular and Developmental Biology – Animal. Click on the title to view the full article at Springerlink.

 

  • Effects of carbendazim and astaxanthin co-treatment on the proliferation of MCF-7 breast cancer cells
    on December 13, 2018 at 12:00 am

    Abstract There has been a controversy in the oncology field about the use of antioxidants along with chemotherapeutics in cancer treatment. This study aimed to investigate the effects of a potent antioxidant (astaxanthin) co-treatment with a promising anti-cancer drug (carbendazim), which is in phase I clinical trials, on MCF-7 breast cancer cell proliferation. MCF-7 cells were treated with carbendazim, astaxanthin, or their combinations and incubated for 24 h. After the incubation, each treatment group was evaluated for proliferation, cell cycle progression, and production of reactive oxygen species (ROS) using WST-1, flow cytometry, and CM-H2DCFDA, respectively. All tested carbendazim and astaxanthin combinations increased the anti-proliferative effect of Carb treatment alone and increased the G2/M phase cell cycle arrest compared to the DMSO-treated control. Astaxanthin, at all concentrations tested, reduced the elevated intracellular ROS levels induced by […]

  • Pterostilbene exerts a protective effect via regulating tunicamycin-induced endoplasmic reticulum stress in mouse preimplantation embryos
    on December 13, 2018 at 12:00 am

    Abstract Pterostilbene (PTS) mainly enriched in small fruits such as berries and grapes exerts an antioxidant effect. However, the protective effects of PTS against endoplasmic reticulum stress (ERS) have not yet been elucidated in mouse preimplantation embryo. ERS plays an important role in regulating the pathological and physiological processes, including embryonic development. We explored the protective effect of PTS on the tunicamycin (TM)-induced ERS in mouse preimplantation embryos. In vitro, culture medium was supplemented with different concentrations of TM and PTS. Our result indicated that treatment of zygotes with 0.5 μg/ml TM significantly decreased the development of day 4 blastocysts (P < 0.05), whereas 0.25 μM PTS supplementation improved the development rate of blastocysts. Moreover, TM treatment significantly increased (P < 0.05) the apoptotic index and reduced the total cell number of the blastocyst, […]

  • Effect of Jieduquyuziyin prescription-treated rat serum on MeCP2 gene expression in Jurkat T cells
    on December 1, 2018 at 12:00 am

    Abstract How genomic DNA methylation and methyl CpG-binding protein 2 (MeCP2) gene expression affect the pathogenesis of systemic lupus erythematosus (SLE) remains poorly understood. Traditional Chinese medicine has a unique effect in the treatment of SLE patients. This study aimed to investigate the effect of Jieduquyuziyin prescription (JP)-treated rat serum on the gene expression of MeCP2 in Jurkat T cells and its role in the pathogenesis of SLE. Jurkat T cells were harvested, and drug-containing serum was prepared. The ferulic acid and paeoniflorin content in the drug-containing serum were determined by liquid chromatography-mass spectrometry (LC-MS/MS). 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assays were used to screen the optimal concentration of drug-containing serum. The DNA methylation level in Jurkat T cells was detected with a Methylamp™ Total DNA Methylation Kit. The methylation status of the […]

  • In vitro culture supplementation of EGF for improving the survival of equine preantral follicles
    on December 1, 2018 at 12:00 am

    Abstract Folliculogenesis is a process of development and maturation of the ovarian follicles, being essential for the maintenance of fertility. In in vivo conditions, 99.9% of the follicles of an ovary do not ovulate and undergo atresia. In order to minimize this loss and to clarify the existing mechanisms, a technique was developed that allows for the in vitro follicular development. The objective of this study was to evaluate the effects of different epidermal growth factor (EGF) concentrations on the in vitro culturing of equine preantral follicles. Ovaries (n = 10) were collected from a local slaughterhouse of mares in seasonal anestrus, washed with 70% alcohol and PBS, and transported. The inner portion of the ovary was divided into 11 fragments of approximately 3 × 3 × 1 mm. A fragment of each ovary was immediately fixed in Bouin (control group). The remaining 10 fragments were individually cultured […]

  • Suppression of STAT3 by S31-201 to reduce the production of immunoinhibitory cytokines in a HIF1-α-dependent manner: a study on the MCF-7 cell line
    on December 1, 2018 at 12:00 am

    Abstract Signal transducer and activator of transcription 3 (STAT3) interacts with many gene promoters and transcription factors such as hypoxia-induced factor 1α (HIF-1α). Recent evidences proposed that STAT3 and HIF-1α together are responsible for angiogenesis and immune response suppression. The main aim of this study was to inhibit STAT3 and HIF-1α and assess their effects on the expression of immunosuppressive cytokines. S31-201 and PX-478 were used to inhibit STAT3 and HIF-1α, respectively. In both hypoxic and normoxic conditions, intracellular levels of HIF-1α were evaluated by western blotting and flow cytometry. Supernatant levels were also measured for VEGF, IL-10, and TGF-β concentration. S31-201 suppressed proliferation of MCF-7 cells and led to reduced HIF-1α expression in both hypoxic and normoxic conditions. It also decreased production of the immunosuppressive cytokines. STAT3 inhibition suppressed […]

  • Establishment of a novel patient-derived Ewing’s sarcoma cell line, NCC-ES1-C1
    on December 1, 2018 at 12:00 am

    Abstract Ewing’s sarcoma is an aggressive mesenchymal tumor characterized by the presence of a unique EWSR1-FLI1 translocation. Ewing’s sarcoma primarily occurs in the bone and soft tissues. Cell lines enable researchers to investigate the molecular backgrounds of disease and the significance of genetic alterations in relevant cellular contexts. Here, we report the establishment and characterization of a novel Ewing’s sarcoma cell line following primary Ewing’s sarcoma tumor tissue culture. The established cell line was authenticated by DNA microsatellite short tandem repeat analysis, characterized by in vitro assays, and named NCC-ES1-C1. The NCC-ES1-C1 cell line grew well for 15 mo and was subcultured more than 50 times during this period. Characterization of the cells revealed that they were not adherent and showed floating features. In conclusion, we successfully established a novel Ewing’s sarcoma cell line, NCC-ES1-C1, from […]

  • Characterization of cell lines derived from the southern armyworm, Spodoptera eridania
    on December 1, 2018 at 12:00 am

    Abstract Spodoptera eridania (southern armyworm) is a polyphagous pest of many plants, including field crops, vegetables, fruits, and ornamentals. Larvae are leaf feeders, defoliating many crops in the tropics and subtropics of the western hemisphere. In this study, cell lines from S. eridania were established to support research focused on the development of advanced pest management technologies. We generated seven cell lines from larval tissues: three from nervous tissues, two from testes, and two from fat bodies. These cell lines have been passaged 18–57 times, indicating they are established lines. They are maintained in EX-CELL 420 or a combination of L15 + EX-CELL 420 media. The identities of the cell lines were confirmed by DAF-PCR and their doubling times ranged from 42 to 110 h. Microscopy indicated the presence of one or more morphologically distinct cell types in each cell line. We identified a catalase gene in all seven cell lines. H2O2 […]

  • Chick embryonic cells as a source for generating in vitro model of muscle cell dystrophy
    on December 1, 2018 at 12:00 am

    Abstract Chick embryonic cells can be used to develop an easy and economical in vitro model for conducting studies on the disease muscle dystrophy (MD). For this, the limb myoblasts from 11th day chick embryo were isolated and cultured. To this muscle cell culture, anti-dystroglycan antibody (IIH6) was added so as to target the α-dystroglycan and disrupt the connection between the cytoskeletal proteins and the extracellular matrix (which is a characteristic feature of MD). Cells were allowed to differentiate further and the morphometrics and mRNA expression were studied. The IIH6-treated muscle cells displayed changes in morphometry, contractibility, and also atrophy was observed when compared to the control cultures. Concomitant gene expression studies showed an upregulation in TGF-β expression, while the muscle sculpture genes MYOD1, MYF5, LAMA2 and MYOG were downregulated resembling the MD in vivo. This simple and cost-effective method can be useful […]

  • Short-term in vitro culture of purity and highly functional rat bone marrow-derived mast cells
    on December 1, 2018 at 12:00 am

    Abstract Mast cells (MCs) are responsible for the innate immune response. Rat MCs are more suitable than mouse MCs as models of specific parasite infection processes and ovalbumin-induced asthma. Rat peritoneum-derived MCs and RBL-2H3 cells (an MC cell line) are widely used in disease studies. However, the application of rat bone marrow-derived MCs (BMMCs) are poorly documented in terms of the methodology of rat BMMC isolation. Here, we describe a relatively rapid, efficient, and simple method for the cultivation of rat BMMCs. As compared to previous protocols, rat BMMCs produced with the proposed protocol exhibited advantages in differentiation, proliferation, lifespan, and functionality, which should prove useful for studies of mucosal MC diseases in specific rat models. […]

  • MiR-126-3p promotes the cell proliferation and inhibits the cell apoptosis by targeting TSC1 in the porcine granulosa cells
    on December 1, 2018 at 12:00 am

    Abstract In mammalian ovaries, many studies demonstrated that the proliferation and apoptosis of granulosa cells are involved in folliculogenesis. Previous evidence suggests that miR-126-3p might get involved in the proliferation and apoptosis of granulosa cells, and tuberous sclerosis complex 1 (TSC1) gene was predicted as one target of miR-126-3p, and moreover, granulosa cell-specific TSC1 knockout stimulated folliculogenesis in mice. However, the molecular regulation of miR-126-3p on TSC1 and its effects on cell proliferation and apoptosis remain virtually unexplored in granulosa cells. Using porcine granulosa cells as a model, the luciferase report assay, mutation, deletion, Annexin-V/PI staining, and EdU assays were applied to investigate the molecular mechanism for miR-126-3p regulating the expression of TSC1 and their effects on the cell proliferation and apoptosis. We found that miR-126-3p showed a positive effect on cell proliferation and a negative effect […]

  • Monensin induces cell death by autophagy and inhibits matrix metalloproteinase 7 (MMP7) in UOK146 renal cell carcinoma cell line
    on December 1, 2018 at 12:00 am

    Abstract Monensin is a metal ionophore used as anticancer agent in many types of cancer cells. In this study, therapeutic potential of monensin was evaluated in TFE3 translocated renal cell carcinoma (RCC) cell line UOK146. UOK146 cells were treated with different concentrations of monensin, and cell death was induced as shown by MTT assay. Autophagy was studied by LC3 western, FACS and LC3 puncta formation after monensin treatment. Mitochondrial potential was studied by staining with TMRM and FACS. Antimetastatic potential of monensin was checked by inhibition of wound closure and MMP7 expression at RNA level. Dead and floating cells after the 10 μM monensin treatment were observed under phase contrast microscope. FACS analysis following TMRM staining showed that mitochondrial membrane gets depolarized after monensin treatment. FACS analysis after acridine orange staining showed increased double positive (green and red) cells, and LC3 upregulation and […]

  • Establishment, characterization, and toxicological application of a spontaneous immortalized cell line from the striped field mouse, Apodemus agrarius
    on December 1, 2018 at 12:00 am

    Abstract It is important to secure various biological resources in situations of diminishing wildlife genetic diversity. Cultured cells are useful bioresources because they can stably store genetic information for a long time and can be expanded efficiently. Here, we established fibroblast cell lines from Apodemus agrarius as a new living resource. A. agrarius is an important sub-predator species in ecosystem food chains and for the study of infection epidemiology. Established cell lines were characterized by chromosome and mitochondrial gene analysis, the observation of cell morphology, and their anchorage-dependent growth pattern. We also examined susceptibility to endocrine disruptors (EDCs), which threaten biodiversity, using these established cell lines. Nonylphenol (NP) is a well-known EDC that threatens wildlife; however, its impact is poorly understood. Sensitivity to NP was confirmed based on two cell viability assays, namely MTT and lactate dehydrogenase. […]

  • CXCR5 overexpression in HL-60 cells enhances chemotaxis toward CXCL13 without anticipated interaction partners or enhanced MAPK signaling
    on December 1, 2018 at 12:00 am

    Abstract CXCR5 is a serpentine receptor implicated in cell migration in lymphocytes and differentiation in leukocytes. It causes MAPK pathway activation and has known membrane partners for signaling. CXCR5 mRNA is reportedly expressed in neutrophils following isolation, but its role in this cellular context is unknown. CXCR5 is also expressed in HL-60 cells, a human acute myeloid leukemia line, following treatment with all-trans retinoic acid, which induces differentiation toward a neutrophil-like state. CXCR5 is necessary for this process; differentiation was crippled in CXCR5 knockout cells and enhanced in cells ectopically expressing it. Since CXCR5 has various membrane protein partners, we investigated whether CXCR5-driven all-trans retinoic acid-induced differentiation depends on its association with such partners. Pursuing this, we generated HL-60 cells overexpressing the protein. We found that CXCR5 drove migration toward its ligand, CXCL13, and probed for […]

  • A novel long non-coding RNA, lncKBTBD10, involved in bovine skeletal muscle myogenesis
    on November 21, 2018 at 12:00 am

    Abstract Accumulating evidence suggests that long non-coding RNAs (lncRNAs) play a crucial role in regulating skeletal muscle myogenesis, a highly coordinated multistep biological process. However, most studies of lncRNAs have focused on humans, mouse, and other model animals. In this study, we identified a novel lncRNA, named lncKBTBD10, located in the nucleus and involved in the proliferation and differentiation of bovine skeletal muscle satellite cells. Prediction of coding potential and in vitro translation system illustrated that lncKBTBD10 has no encoding capability. With the process of myogenic differentiation, the expression of lncKBTBD10 gradually increased. To elucidate the functions of lncKBTBD10 during myogenesis, the gain/loss-of-function experiments were performed. Results showed that the proliferation and differentiation of bovine skeletal muscle satellite cells were all suppressed whether lncKBTBD10 was knocked down or over-expressed. Furthermore, we […]

  • Butyric acid induces spontaneous adipocytic differentiation of porcine bone marrow–derived mesenchymal stem cells
    on November 19, 2018 at 12:00 am

    Abstract Butyric acid (BA) affects the differentiation of mesenchymal stem cells (MSC) through the activation of different transcriptional pathways. The aim of this study was to determine the effects of BA on proliferation and spontaneous differentiation of porcine bone marrow–derived MSC. Second passage MSC (n = 6) were cultured in either a basal medium (BM, DMEM + 10% FBS), or BM + 2.5 mmol/L BA (BA-2.5) or BM + 5 mmol/L BA (BA-5). Cell proliferation was significantly decreased by both BA-2.5 and BA-5 after 48 h and 72 h (− 55% and − 63%, respectively). To assess the impact of BA on spontaneous differentiation, MSC were cultured for 27 d, with complete media changes every 3 d. At day 27, cells were stained for osteocytic, chondrocytic, and adipocytic differentiation. No terminal differentiation was detected in control MSC, while accumulated small […]

  • Role of miR-9-5p in preventing peripheral neuropathy in patients with rheumatoid arthritis by targeting REST/miR-132 pathway
    on November 19, 2018 at 12:00 am

    Abstract MicroRNAs (miRNAs) are found to play a key role in neural cell differentiation, peripheral nerve injury, and rheumatoid arthritis (RA). However, no study has yet been conducted highlighting their role in RA-induced peripheral neuropathy. Here, we investigated the role of miRNAs in RA-induced peripheral neuropathy. Levels of six miRNAs were detected in serum collected from 15 patients with RA and peripheral neuropathy and 16 patients with RA. In vitro, Schwann cells were treated with 0.1 ng/mL IL-6 and 20 ng/mL TNF-α. The expression level of miR-9-5p and its association with the repressor element-1 silencing transcription factor (REST) were investigated. The roles of miR-9-5p and REST in Schwann cell injury were examined after transfection of miR-9-5p mimics or REST siRNA. In patients with RA and peripheral neuropathy, serum miR-9-5p was significantly downregulated when compared with RA. In IL-6- and TNF-α-stimulated Schwann cells, […]

  • Improved synergistic anticancer efficacy of quercetin in combination with PI-103, rottlerin, and G0 6983 against MCF-7 and RAW 264.7 cells
    on November 9, 2018 at 12:00 am

    Abstract Flavonoids have been chronicles of the history of a long way journey in the cure of physiological or pathophysiological conditions in various diseases including cancer. Our previous findings suggest the extensive mechanism of quercetin (QUE) mediated regression of cell survival, cell proliferation, oxidative stress, inflammation, and angiogenesis via modulating PI3K and PKC signaling in lymphoma as well as hepatocellular carcinoma. PI3K-PKC pathway is a key monitor of mammalian cells regulated by its different isoenzymes, which may exert similar or opposite cellular effects by differential coupling of signaling pathways. Put forward the invention of selective inhibitors against various isoenzymes is beneficial to reduce the burden of inclusive deleterious effects of drug for normal physiological process. Therefore, we hypothesized the improved anticancer efficacy of QUE in combination with isoenzyme inhibitors—rottlerin (ROT-PKCδ inhibitor), G0 […]

  • Establishment of novel patient-derived models of dermatofibrosarcoma protuberans: two cell lines, NCC-DFSP1-C1 and NCC-DFSP2-C1
    on November 8, 2018 at 12:00 am

    Abstract Dermatofibrosarcoma protuberans (DFSP) is a common type of dermal sarcoma, characterized by the presence of the unique collagen type I alpha 1 chain (COL1A1)-PDGFB translocation, which causes constitutive activation of the platelet-derived growth factor β (PDGFB) signaling pathway. Patients with DFSP exhibit frequent local recurrence, and novel therapeutic approaches are required to achieve better clinical outcomes. Patient-derived cancer cell lines are essential in the preclinical research. Here, we established novel patient-derived DFSP cell lines from two patients with DFSP and designated these cell lines NCC-DFSP1-C1 and NCC-DFSP2-C1. Tumors of the two patients with DFSP had COL1A1-PDGFB translocations with distinct COL1A1 breakpoints, e.g., in exons 33 and 15, and the translocations were preserved in the established cell lines. NCC-DFSP1-C1 and NCC-DFSP2-C1 cells exhibited similar morphology and limited capability of proliferation in vitro, forming […]

  • Production of 8-nitro-cGMP in osteocytic cells and its upregulation by parathyroid hormone and prostaglandin E 2
    on November 5, 2018 at 12:00 am

    Abstract Osteocytes regulate bone remodeling, especially in response to mechanical loading and unloading of bone, with nitric oxide reported to play an important role in that process. In the present study, we found that 8-nitroguanosine 3′,5′-cyclic monophosphate (8-nitro-cGMP), a second messenger of nitric oxide in various types of cells, was produced by osteocytes in bone tissue as well as cultured osteocytic Ocy454 cells. The amount of 8-nitro-cGMP in Ocy454 cells increased during incubation with parathyroid hormone or prostaglandin E2, both of which are known to upregulate receptor activator of nuclear factor-κB ligand (RANKL) mRNA expression in osteocytes. On the other hand, exogenous 8-nitro-cGMP did not have effects on either the presence or absence of these bioactive substances. Furthermore, neither an inhibitor of nitric oxide synthase nor 8-bromo-cGMP, a cell-permeable analog of cGMP, showed remarkable effects on mRNA expression of […]

  • Antitumor effects of SB injection in canine osteosarcoma and melanoma cell lines
    on October 31, 2018 at 12:00 am

    Abstract The present study was designed to evaluate the effect of SB injection, which is composed of extracts from the roots of Pulsatilla koreana, Panax ginseng, and Glycyrrhiza glabra, on the viability of canine osteosarcoma and melanoma cells and nonneoplastic canine cells. Cells were treated with SB injection, conventional chemotherapeutic drugs, or a combination of both at various concentrations. Cellular viability was evaluated using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Flow cytometry was used to evaluate the cell cycle and apoptosis. SB injection inhibited the growth of osteosarcoma and melanoma cells in a dose-dependent manner. The cell cycle of the affected cells was arrested in the G2/M phase, indicating an anti-proliferative effect. SB injection dose-dependently increased the rate of apoptosis. Furthermore, we found that combining SB injection with chemotherapeutic drugs resulted in a greater reduction in canine […]



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