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.

 

  • Repression of insulin gene transcription by indirect genomic signaling via the estrogen receptor in pancreatic beta cells
    on February 21, 2019 at 12:00 am

    Abstract The mechanism whereby 17β-estradiol (E2) mediates insulin gene transcription has not been fully elucidated. In this study, exposure of hamster insulinoma (HIT-T15) cells to 5 × 10−9 to 1 × 10−7 M E2 led to a concentration-dependent decrease of insulin mRNA levels. Transient expression of the estrogen receptor (ER) in HIT-T15 cells revealed that estrogen receptor α (ERα) repressed transcription of the rat insulin II promoter in both ligand-dependent and ligand-independent manners. The N-terminal A/B domain of ERα was not required for either activity. However, the repression was absent with mutated ER lacking the DNA-binding domain. Moreover, introducing mutations in the D-box and P-box of the zinc finger of ER (C227S, C202L) also abolished the repression. Deletion of the insulin promoter region revealed that nucleotide positions − 238 to − 144 (relative […]

  • Exosomes derived from mesenchymal stem cells inhibit mitochondrial dysfunction-induced apoptosis of chondrocytes via p38, ERK, and Akt pathways
    on February 19, 2019 at 12:00 am

    Abstract Osteoarthritis (OA) is the most common chronic joint disease worldwide. Chondrocyte, as the only resident cell type in cartilage, its apoptosis is of pathogenetic significance in OA. Mesenchymal stem cell (MSC)-based-therapy has been proved effective in OA in animals and clinical studies. Nowadays, the regenerative potential of MSC-based therapy is mostly attributed to its paracrine secretion, in which exosomes may play an important role. In the present study, we aimed to find out the significance of MSC-derived exosomes (MSC-Exos) on the viability of chondrocytes under normal and inflammatory conditions. Bone marrow MSCs (BMSCs) and chondrocytes from rabbits were cultured in vitro. BMSC-Exos were isolated by an ultracentrifugation method. Transmission electron microscopy and Western blot were used to identify exosomes. The internalization of BMSC-Exos into chondrocytes was observed by fluorescent microscope. The viability and apoptosis of chondrocytes […]

  • Effect of attenuation of fibroblast growth factor receptor 2b signaling on odontoblast differentiation and dentin formation
    on February 12, 2019 at 12:00 am

    Abstract Attenuation of fibroblast growth factor receptor (FGFR) 2b signaling suppresses the differentiation of oral epithelial stem cells to ameloblasts, their survival and viability remaining unaffected; however, its effect on dentin formation is unknown. This study aimed to clarify the effect of attenuation of FGFR2b signaling on odontoblast differentiation and dentin formation. Initially, we used a murine rtTA transactivator/tetracycline promoter system for inducible and reversible attenuation of FGFR2b signaling in adult mice. Experimental animals overexpressed soluble FGFR2b (sFGFR2b), and wild-type controls were selected from the same litter (WT group). Histological analysis of CMV mice confirmed the obliteration of the enamel and ameloblast layer, and micro CT analysis revealed a significant increase in dentin thickness in CMV mice rather than in WT mice (P < 0.05). On analyzing the expression of dentin-related differentiation factors, DSPP, […]

  • Genetic algorithm as an optimization tool for the development of sponge cell culture media
    on February 11, 2019 at 12:00 am

    Abstract Sponges are rich sources of novel natural products. Production in cell cultures may be an option for supply of these compounds but there are currently no sponge cell lines. Because there is a lack of understanding about the precise conditions and nutritional requirements that are necessary to sustain sponge cells in vitro, there has yet to be a defined, sponge-specific nutrient medium. This study utilized a genetic algorithm approach to optimize the amino acid composition of a commercially available basal cell culture medium in order to increase the metabolic activity of cells of the marine sponge Dysidea etheria. Four generations of the algorithm were carried out in vitro in wet lab conditions and an optimal medium combination was selected for further evaluation. When compared to the basal medium control, there was a twofold increase in metabolic activity. The genetic algorithm approach can be used to optimize other components of culture media to […]

  • miR-145-5p suppresses osteogenic differentiation of adipose-derived stem cells by targeting semaphorin 3A
    on February 11, 2019 at 12:00 am

    Abstract Adipose-derived stem cells (ADSCs) provide a novel method for bone tissue regeneration, but their adipogenic tendency limits their therapeutic efficacy. MicroRNAs (miRNAs) have been reported to regulate stem cell differentiation and bone tissue regeneration, but the detailed mechanism is poorly investigated. Our study indicated that inhibition of miR-145-5p enhanced the osteogenic potential of ADSCs and reduced the adipogenic differentiation. Osteogenesis- and adipogenesis-associated genes were detected by qRT-PCR indicating a corresponding result. Moreover, semaphorin 3A (sema3A) was found to be a target of miR-145-5p, as confirmed by a luciferase activity assay, qRT-PCR, and western blotting. Inhibition of miR-145-5p promoted migration, as detected by wound healing and Transwell assays, but did not affect proliferation, as detected by CCK-8 and ki-67 assays. The effects of miR-145-5p inhibitors on ADSC progression rescued by siRNA of Sema3a and si-sema3a […]

  • Calcium ionophore enhanced developmental competence and apoptotic dynamics of goat parthenogenetic embryos produced in vitro
    on February 8, 2019 at 12:00 am

    Abstract Parthenogenetically developed embryos are efficient sources of in vitro embryo production, having less ethical issue and being useful for investigating culture conditions/treatments, early developmental, genomic studies, and homonymous source of stem cells. Keeping its advantages in mind, we aimed to study the effects of different activating agents on embryo production and its quality and gene expression. In the present study, 1348 immature oocytes recovered were parthenogenetically developed to embryos. Usable-quality immature oocytes were collected by puncturing the surface follicles and matured in in vitro maturation (IVM) medium for 27 h in a humidified 5% CO2 incubator at 38.5°C. The matured oocytes were parthenogenetically activated by exposure to 5 μM calcium ionophore for 5 min or 7% ethanol for 7 min sequentially followed by 4 h incubation in 2 mM 6-DMAP and then in vitro cultured (IVC) in RVCL/G-2 medium for […]

  • Retinoic acid (RA) and bone morphogenetic protein 4 (BMP4) restore the germline competence of in vitro cultured chicken blastodermal cells
    on February 8, 2019 at 12:00 am

    Abstract Chicken blastodermal cells (BCs) are pluripotent stem cells derived from early embryos and may be easily obtained and manipulated. However, in vitro cultured BCs have extremely low germline capacity, which may limit their applications. Research on the germ cell differentiation of mammalian pluripotent cells using chemical-inducing agents has gained popularity, and tremendous achievements have been made. Whether chemical-inducing agents allow acquirement of germline competence in BCs is, however, questionable. In this study, retinoic acid (RA) and bone morphogenetic protein 4 (BMP4) promoted the expression of germline-specific genes and restored the germline competence of in vitro cultured BCs. Moreover, BCs induced with RA and BMP4 could efficiently produce gonadal chimeric chick embryos. These results may greatly enhance the potential applications of BCs in biotechnology and basic research. […]

  • Establishment of an electroporation-mediated gene delivery system in porcine spermatogonial stem cells
    on February 6, 2019 at 12:00 am

    Abstract Spermatogonial stem cells (SSCs) are a useful tool for the generation of genetically modified transgenic sperm. As a result, the transfer of specific genes into the cytoplasm of SSCs is crucial for the successful generation of transgenic sperm. Here, we report electroporation conditions optimized for SSCs derived from the porcine testis. The highest transfection efficiency and cell viability were observed in porcine SSCs transfected with 1 μg transgenic vector with a single electric pulse from an electroporator at a voltage of 100 V and a capacitor setting of 250 μF. The transfection efficiency and cell viability were constant regardless of the size of the transgenic vector. Furthermore, we did not detect loss of spermatozoa differentiation potential in the transfected porcine SSCs. From these results, we confirm that this electroporation-based gene delivery system can effectively introduce foreign DNA into the genome of porcine SSCs […]

  • Effects of carbendazim and astaxanthin co-treatment on the proliferation of MCF-7 breast cancer cells
    on February 1, 2019 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 February 1, 2019 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 […]

  • Conditions facilitating infection of mosquito cell lines with Wolbachia , an obligate intracellular bacterium
    on February 1, 2019 at 12:00 am

    Abstract Factors that influence establishment of Wolbachia, an obligate intracellular bacterium, in novel insect hosts or uninfected insect cell lines are poorly understood. Infectivity of Wolbachia strain wStr was correlated with flow cytometric profiles to define optimal conditions for harvesting an infectious inoculum. Wolbachia recovered from the cell culture supernatant after gentle pipetting of infected cells represented about 1% of the total bacterial population and were more infectious than Wolbachia that remained associated with intact cells and/or membranes after low-speed centrifugation. Optimal establishment of a robust infection in naïve cells required 6 d, at a ratio of 80 to 160 bacteria per cell. Among Aedes albopictus mosquito cell lines, an aneuploid line with a 4n + 1 karyotype was more susceptible to infection than diploid lines. These findings contribute to the in vitro manipulation of Wolbachia, illustrate some of the […]

  • Gene delivery into Siberian sturgeon cell lines by commercial transfection reagents
    on February 1, 2019 at 12:00 am

    Abstract The optimal transfection conditions for efficient transgene delivery into a specific cell type should be empirically determined, particularly in cases involving unusual cell types. We compared the conditions for effective introduction of transgenes into Siberian sturgeon (Acipenser baerii) cell lines by evaluating the cytotoxicity and transfection efficiency of three commercially available transfection reagents: Lipofectamine 2000, X-tremeGENE HP DNA Transfection Reagent, and GeneJuice Transfection Reagent. Plasmid vectors containing the gene encoding enhanced green fluorescent protein were mixed with each of the transfection reagents using reagent-to-plasmid ratios of 1:1, 2:1, and 4:1. Then, the complexes were used to transfect three Siberian sturgeon cell lines derived from the heart, head kidney, and gonad. Cytotoxicity and transfection efficiency were measured via flow cytometry after propidium iodide staining. No significant cytotoxicity was observed […]

  • Human menstrual blood–derived stem cells protect H9c2 cells against hydrogen peroxide–associated apoptosis
    on February 1, 2019 at 12:00 am

    Abstract Human menstrual blood–derived mesenchymal stem cells (MenSCs) hold great promise for regenerative medicine. Here, H2O2-associated damage in H9c2 cells was employed as an in vitro ischemia–reperfusion model, and the transwell system was used to explore the beneficial effects of MenSCs on the H2O2-induced damage of myocardial H9c2 cells. H2O2 treatment resulted in decreased viability and migration rate, with increased apoptosis levels in cells. By contrast, upon co-culture with MenSCs, H9c2 cell viability and migration were increased, whereas the apoptotic rate decreased. Additionally, western blot and qRT-PCR showed that MenSCs mediated the anti-apoptotic role by downregulating the pro-apoptotic genes Bax and caspase-3, while upregulating the anti-apoptotic effector Bcl-2. Furthermore, co-culture with MenSCs resulted in elevated expression of N-cadherin after H2O2 treatment. These findings indicate that MenSCs protect H9c2 cells against […]

  • Renogenic characterization and in vitro differentiation of rat amniotic fluid stem cells into renal proximal tubular- and juxtaglomerular-like cells
    on February 1, 2019 at 12:00 am

    Abstract The aim of the present study was to investigate the renogenic characteristics of amniotic fluid stem cells (AFSCs) and to evaluate their in vitro differentiation potential into renal proximal tubular-like cells and juxtaglomerular-like cells. We culture expanded AFSCs derived from rat amniotic fluid. The AFSCs grew as adherent spindle-shaped cells and expressed mesenchymal markers CD73, CD90, and CD105 as well as renal progenitor markers WT1, PAX2, SIX2, SALL1, and CITED1. AFSCs exhibited an in vitro differentiation potential into renal proximal tubular epithelial-like cells, as shown by the upregulation of expression of proximal tubular cell–specific genes like AQP1, CD13, PEPT1, GLUT5, OAT1, and OCT1. AFSCs could also be differentiated into juxtaglomerular-like cells as demonstrated by the expression of renin and α-SMA. The AFSCs also expressed pluripotency markers OCT4, NANOG, and SOX2 and could be induced into embryoid bodies with […]

  • 2018 Special Reviewers Thank You
    on February 1, 2019 at 12:00 am

    […]

  • Association of Gpx1 fluctuation in cell cycle progression
    on February 1, 2019 at 12:00 am

    Abstract This research demonstrates fluctuation of glutathione peroxidase1 (Gpx1) throughout cell cycle progression with significant decreased expression at mitosis of HeLa cell. This was achieved with western blot (WB) analysis of target proteins from each phase of synchronized cells. The synchronizations were performed with double thymidine (T/T) for G1/S arrest and thymidine followed by nocodazole (T/N) for G2/M arrest. The G1/S arrested cells were released in fresh medium for 3, 6, 9, 10, and 15h to obtain cell at each phase such as gap1 (G1), synthesis (S), gap2 (G2), mitosis (M), and gap1 (G1) phase, respectively, for investigating Gpx1 expression throughout a complete cycle. The synchronizations were confirmed using fluorescence activated cell sorting (FACS) and WB analysis of phase-specific markers. The fluctuations of Gpx1 expression were verified with universal protein actin and peroxiredoxin1 (Prx1) which are stable throughout the cell cycle. […]

  • Differentiation of mouse-induced pluripotent stem cells into dental epithelial-like cells in the absence of added serum
    on February 1, 2019 at 12:00 am

    Abstract Recent studies have successfully generated tooth-like structure by mimicking the reciprocal interaction between dental epithelial and mesenchymal cells in tooth organogenesis. However, clinical applications of these methods are limited primarily due to the lack of appropriate sources for dental epithelial cells. Induced pluripotent stem cells (iPSCs) are attractive as a source for dental epithelial cells due to their unique characteristics. In this study, we examined the effect of neurotrophin-4 (NT-4) on the differentiation of mouse iPSCs (miPSCs) into dental epithelial cells. Our results showed that the addition of NT-4 during the formation of embryoid body significantly triggered the upregulation of epithelial markers such as p63 and CK14, suggesting that NT-4 provides an inductive condition for the differentiation of miPSCs into epithelial cells. Expansion of the NT-4-treated cells under serum-free culture conditions improves the formation of cells […]

  • Production of 8-nitro-cGMP in osteocytic cells and its upregulation by parathyroid hormone and prostaglandin E 2
    on January 1, 2019 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 […]

  • Butyric acid induces spontaneous adipocytic differentiation of porcine bone marrow–derived mesenchymal stem cells
    on January 1, 2019 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 […]

  • A novel long non-coding RNA, lncKBTBD10, involved in bovine skeletal muscle myogenesis
    on January 1, 2019 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, […]


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