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.

 

  • Differential effects of extracellular matrix proteins on in vitro culture and growth characteristics of caprine male germ cells
    on April 7, 2021 at 12:00 am
  • Synergic effects of extremely low-frequency electromagnetic field and betaine on in vitro osteogenic differentiation of human adipose tissue-derived mesenchymal stem cells
    on March 26, 2021 at 12:00 am

    Abstract Human adipose tissue-derived mesenchymal stem cells (hADSCs) due to easy extraction, relative abundance, in vitro expansion and differentiation potential, frozen storage capability, and ability to secrete cytokines, compared to other stem cells, are appropriate candidate in regenerative medicine. Extremely low-frequency electromagnetic fields (ELF-EMF) and betaine are two safe factors in bone lesions repair. This study was designed to assess the osteogenic differentiation potential of these factors on hADSCs. The samples were collected from women undergoing liposuction after obtaining written consent. The hADSCs were extracted and treated with osteogenesis differentiation medium (OD) as the positive control, with OD and betaine (BET group), with OD and EMF (EMF group), and with OD and betaine and EMF (BET+EMF group) for 21 d; the negative control consisted of cells without treatment. Betaine 10 mM and EMF with 50-Hz frequency, 1-mT intensity (8 h daily), and […]

  • 3,5-Dimethoxy-4-benzoic acid (syringic acid) a natural phenolic acid reduces reactive oxygen species in differentiated 3T3-L1 adipocytes
    on March 26, 2021 at 12:00 am

    Abstract Preadipocytes under nutrient excess mature to lipid-laden adipocytes that are hotspots for generation of reactive oxygen species (ROS) imbalance and oxidative stress. Syringic acid (SA), a natural phenolic acid, was evaluated for its in vitro antioxidant and ROS modulation during in matured 3T3-L1 adipocytes. Following 10 d, the SA-treated adipocytes were evaluated for the levels of glutathione (GSH) and antioxidant enzymes such as superoxide dismutase (SOD) and catalase (CAT). The levels of peroxides in mature adipocytes were estimated using dichlorofluorescein (DCF) cleavage fluorescence. The level of NADPH oxidase 4 (NOX4) expression was also investigated following 10-d differentiation period. SA significantly improved the levels of GSH, SOD, and CAT in matured adipocytes. Reduction in ROS production levels was also witnessed by decrease in DCF cleavage. SA showed concentration-dependent inhibition of NOX4 by day 7 of adipogenesis when compared with […]

  • MiR-375 potentially enhances GnRH expression by targeting Sp1 in GT1-7 cells
    on March 26, 2021 at 12:00 am

    Abstract Gonadotropin-releasing hormone (GnRH) is the initial central regulator of the animal reproduction system, which is crucial for puberty onset and fertility. However, the mechanisms regulating GnRH production and release remain unclear. In addition, few studies reported that miR-375 expressed in mouse hypothalamus, but up to now there are limited functional studies of miR-375 in regulating GnRH secretion. According to our recent findings that miR-375 was involved in regulating the synthesis and secretion of pituitary hormones, thus, we aimed to identify the role of miR-375 in regulating GnRH production in GT1-7 cells. Immunofluorescence results demonstrated that miR-375 was expressed in all of the GT1-7 cells. The functional studies showed that miR-375 overexpression enhanced GnRH mRNA expression level, but decreased the mRNA expressions of Sp1, Cebpb, Msx1, and Tle4. Transcriptomics analysis demonstrated Sp1 and Tle4 acted as the targeting genes of miR-375, […]

  • RGMa can induce skeletal muscle cell hyperplasia via association with neogenin signalling pathway
    on March 21, 2021 at 12:00 am

    Abstract Although originally discovered inducing important biological functions in the nervous system, repulsive guidance molecule a (RGMa) has now been identified as a player in many other processes and diseases, including in myogenesis. RGMa is known to be expressed in skeletal muscle cells, from somites to the adult. Functional in vitro studies have revealed that RGMa overexpression could promote skeletal muscle cell hypertrophy and hyperplasia, as higher efficiency in cell fusion was observed. Here, we extend the potential role of RGMa during C2C12 cell differentiation in vitro. Our results showed that RGMa administrated as a recombinant protein during late stages of C2C12 myogenic differentiation could induce myoblast cell fusion and the downregulation of different myogenic markers, while its administration at early stages induced the expression of myogenic markers with no detectable morphological effects. We also found that RGMa effects on skeletal muscle […]

  • Silencing MAP3K1 expression inhibits the proliferation of goat hair follicle stem cells
    on March 21, 2021 at 12:00 am

    Abstract The Yangtze River Delta White Goat is the only goat breed in the world that can produce superior-quality brush hair. Previous studies have shown that some genes are expressed differentially in the skin tissues between the goats produced superior-quality and normal-quality brush hair. Studies also have shown that different gene play varied roles in regulating the proliferation and apoptosis of hair follicle stem cells. However, the biological function of MAP3K1 (mitogen-activated protein kinase kinase kinase 1) gene in hair follicle stem cells is not fully understood. This study aims to investigate the role of MAP3K1 knockdown during the proliferation and apoptosis of hair follicle stem cells. RT-qPCR and Western blot were used to detect mRNA gene and protein expression level, CCK-8 and EdU assays were used to detect cell proliferation, and cell cycle and apoptosis were detected by flow cytometry. The results showed that the MAP3K1 expression level was […]

  • Icariside II promotes the differentiation of human amniotic mesenchymal stem cells into dopaminergic neuron-like cells
    on March 15, 2021 at 12:00 am

    Abstract The purpose of this study is to observe the effect of icariside II (ICS II) on the differentiation of human amniotic mesenchymal stem cells (hAMSCs) into dopaminergic neuron-like cells, the involvement of PI3K signaling pathway inhibitors. After identifying hAMSCs by flow cytometry, hAMSCs were induced and treated with ICS II at 10 μmol/L, 3 μmol/L, 1 μmol/L, and 0 μmol/L. hAMSCs in the LY294002+3μM ICS II group were pretreated with 20 μmol/L LY294002, a PI3K-specific inhibitor, for 1 h, and then hAMSCs were induced with 3 μmol/L ICS II. On the 21st day of induction, immunofluorescence was used to detect expression of the neuronal nuclei (NeuN), neuron-specific enolase (NSE), microtubule-associated protein-2 (MAP-2), glial fibrillary acidic protein (GFAP), and tyrosine hydroxylase (TH) antigens in each induced cell group. Western blotting was used to detect the relative protein expression of NSE, MAP-2, GFAP, and TH. ELISA was used to detect the […]

  • Involvement of miR-337 in high glucose–suppressed osteogenic differentiation in bone marrow mesenchymal stem cells via negative regulation of Rap1A
    on March 1, 2021 at 12:00 am

    Abstract This study aims to investigate the inhibitory effect of microRNA-337 (miR-337) on osteogenic differentiation in bone marrow mesenchymal stem cells and its action of mechanisms. Overexpression and knockdown of miR-337 were performed in bone marrow mesenchymal stem cells (BMSCs). Cell proliferation was assessed by using a cell counting kit-8 (CCK-8), mineralization assay was performed by alizarin red staining, and alkaline phosphatase activity was then measured. Luciferase reporter assay was applied to verify miR-337 binding to Ras-related protein 1A (Rap1A) mRNA. Reverse transcription and quantitative polymerase chain reaction (RT-qPCR) was applied to measure the expressions of runt-related transcription factor 2 (Runx2), alkaline phosphatase (ALP), osteocalcin (OCN), osteopontin (OPN), bone morphogenetic protein (BMP2), and miR-337. Then the protein level of Rap1A was determined by western blot analysis. High glucose inhibited osteogenic differentiation […]

  • Hormesis of low-dose inhibition of pAkt1 (Ser473) followed by a great increase of proline-rich inositol polyphosphate 5-phosphatase (PIPP) level in oocytes
    on March 1, 2021 at 12:00 am

    Abstract Hormesis describes a biphasic dose-response relationship generally characterized by a low-dose excitement and a high-dose inhibition. This phenomenon has been observed in the regulation of cell, organ, and organismic level. However, hormesis has not reported in oocytes. In this study, we observed, for the first time, hormetic responses of PIPP levels in oocytes by inhibitor of Akt1 or PKCδ. The expression of PIPP was detected by qPCR, immunofluorescent (IF), and Western Blot (WB). To observe the changes of PIPP levels, we used the inhibitors against pAkt1 (Ser473) or PKCδ, SH-6 or sotrastaurin with low and/or high-dose, treated GV oocytes and cultured for 4 h, respectively. The results showed that PIPP expression was significantly enhanced when oocytes were treated with SH-6 or sotrastaurin 10 μM, but decreased with SH-6 or sotrastaurin 100 μM. We also examined the changes of PIPP levels when GV oocytes were treated with exogenous PtdIns(3,4,5)P3 or […]

  • Copper toxicity of inflection point in human intestinal cell line Caco-2 dissected: influence of temporal expression patterns
    on March 1, 2021 at 12:00 am

    Abstract We previously described a non-monotonic dose response curve at low copper concentrations where 3.125 μM CuSO4 (the early inflection point) was more toxic than 25 μM CuSO4 in Caco-2 cells. We employed global proteomics to investigate this observation. The altered expression levels of a small number of proteins displaying a temporal response may provide the best indication of the underlying mechanism; more well-known copper response proteins including the metal binding metallothioneins (MT1X, MT1F, MT2A) and antioxidant response proteins including Heme oxygenase were upregulated to a similar level in both copper concentrations and so are less likely to underpin this phenomenon. The temporal response proteins include Granulins, AN1-type zinc finger protein 2A (ZFAND2A), and the heat shock proteins (HSPA6 and HSPA1B). Granulins were decreased after 4 h only in 25 μM CuSO4 but from 24 h, were decreased in both copper concentrations to a […]

  • Specific protein 1 inhibitor mithramycin A protects cardiomyocytes from myocardial infarction via interacting with PARP
    on March 1, 2021 at 12:00 am

    Abstract Specific protein 1 (SP1) might act as a critical transcription regulator in myocardial infarction (MI), but little evidence about its function in regulating cardiac apoptosis, a major cause of MI development, has been revealed. This study tried to investigate the role of SP1 in MI and its interaction with poly-ADP-ribose polymerase (PARP)-1 by using SP1 inhibitor, mithramycin A (mithA). Primary mouse cardiomyocytes and commercial mouse cardiomyocytes were subjected to mithA treatment under hypoxia conditions, while cell viability, Nix promoter activity, and its expression were detected correspondingly. PARP overexpression and knockdown were conducted, respectively, in mithA-treated and SP1-overexpressing cells. Co-immunoprecipitation was used to verify the interaction between PARP and SP1. For in vivo experiments, mithA administration was performed after the injections of adenovirus for PARP overexpression, and then, MI introduction was carried out. […]

  • MiR-26b-5p regulates the preadipocyte differentiation by targeting FGF 21 in goats
    on March 1, 2021 at 12:00 am

    Abstract MicroRNAs are a class of highly conserved and widely distributed non-coding RNAs. It is known that miR-26b has a high abundance in adipose tissue and is considered to be an effective regulator of adipogenesis. However, it is unclear whether miR-26b-5p, the product of miR-26b precursor, has the same effect as miR-26b. In the present study, we explored the potential role of miR-26b-5p in preadipocyte differentiation of goats. We found that the expression of miR-26b-5p had dramatic change during goat intramuscular preadipocyte differentiation. Transfection and RT-qPCR revealed that overexpression of miR-26b-5p increased the level of adipogenic marker genes and lipid accumulation in goat preadipocyte, suggesting that miR-26b-5p positively regulates goat preadipocyte differentiation. Furthermore, bioinformatics analysis and dual fluorescein reporter assays were performed to predict and validate the targets of miR-26b-5p. The results showed that miR-26b-5p has a […]

  • Stimulation of melanin synthesis by UVB is mediated by NO/cGMP/PKG cascade targeting PAK4 in vitro
    on March 1, 2021 at 12:00 am

    Abstract The coat color of mammals is primarily determined by the type, quantity, and distribution of melanin in the skin and hair. As an endogenous gas molecule, nitric oxide (NO) regulates tyrosinase production by modulating the cGMP-dependent protein kinase (PKG) pathway, which enhances melanin synthesis. However, some interrelationships have not been fully elucidated. In the present study, mouse melanocytes co-cultured with mouse keratinocytes in vitro, or as monocultures, were used as research models. The results indicated that ultraviolet B irradiation increased nitric oxide synthase (NOS) activity and NO production, and increased PKG, p21-activated kinase 4 (PAK4), and microphthalmia-associated transcription factor (MITF) levels, as well as tyrosinase (TYR), tyrosinase-related protein 1 and 2 expression, and melanin synthesis. During PKG inhibition, the expression of NO-regulated PAK4 and MITF was decreased. Pigment production was also affected, but remained […]

  • Dp44mT regulates the levels of inflammatory mediators through blocking NF-κB nuclear translocation in LPS-stimulated RAW 264.7 macrophages
    on March 1, 2021 at 12:00 am

    Abstract Inflammation is increased by infection with pathogens such as viruses, bacteria, and parasites. High levels of inflammatory mediators and infiltration of macrophages into inflammatory lesions were reported in severe inflammatory diseases. Here, the aim of this study was to evaluate an anti-inflammatory activity of di-2-pyridylketone-4,4-dimethyl-3-thiosemicarbazone (Dp44mT) on lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. Dp44mT (1–100 ng/mL) had no effect on viability of RAW 264.7 macrophages. Dp44mT (100 ng/mL) significantly reduced LPS-induced release of nitric oxide and expression of inducible nitric oxide synthase and cyclooxygenase-2. A significant upregulation of tumor necrosis factor (TNF)-α and interleukin (IL)-6 by LPS stimulation was downregulated by treatment with Dp44mT. Dp44mT blocked activation of nuclear factor-κB by the interruption of IκBα phosphorylation. Dp44mT suppressed the phagocytosis. Furthermore, administration […]

  • Bifenthrin induces DNA damage and autophagy in Spodoptera frugiperda (Sf9) insect cells
    on March 1, 2021 at 12:00 am

    Abstract Bifenthrin is one of the most commonly used synthetic pyrethroid insecticides. It targets the nervous system of insects, mainly acting on sodium channels in nerve cell membranes. The high use of bifrenthrin may lead to an increase in pest insect resistance. Additionally, there are only a few studies describing its cytotoxic action. A series of bioassays were carried out, and the results showed that bifenthrin has a significant ability to induce DNA damage and the inhibition of viability in Spodoptera frugiperda (Sf9) cells. Monodansylcadaverine staining and transmission electron microscope assays were used to observe significant levels of autophagosomes and mitochondrial dysfunction in the cytoplasm. Additionally, western blot analysis showed an upregulation in LC3-II and beclin-1 protein expression and a downregulation in p62 expression, which contributed to the cytotoxic effect of bifenthrin on Sf9 cells. Overall, bifenthrin significantly impacts the […]

  • Hyperosmolarity benefits cartilage regeneration by enhancing expression of chondrogenic markers and reducing inflammatory markers
    on March 1, 2021 at 12:00 am

    Abstract Application of hyperosmolarity can be a promising strategy to promote chondrogenic differentiation in adipose-derived mesenchymal stem cells (ADSCs). Growth factors may promote different signaling pathways in parallel that is why in this study we monitor undesired pathologic or unwanted side effects as well as chondroinductive impacts of hyperosmolarity in differentiating ADSCs. Quantified gene expression, immunocytochemistry, glycosaminoglycan deposition and angiogenic secretion assays performed along with immunoassay. We observed that hyperosmolarity pressure of 480 mOsm over-expressed cartilage specific markers at gene expression level in the extra cellular matrix. Meanwhile, hyperosmolarity of 480 mOsm diminished the expression of cartilage associated pathologic markers, i.e., inflammatory and angiogenic attributes. Certain dose of hyperosmolarity could benefit chondrogenesis in a dual way, first by increasing chondrogenic markers and second by […]

  • MiR-23b-3p functions as a positive factor for osteoporosis progression by targeting CCND1 in MC3T3-E1 cells
    on March 1, 2021 at 12:00 am

    Abstract MiRNAs have gained tremendous attention as studies have shown that miRNAs play important roles in osteoporosis (OP) progression. This study attempted to explore whether miR-23b-3p is involved in the pathogenesis of OP. We detected the miR-23b-3p and Cyclin D1 (CCND1) expressional patterns in the bone of patients with or without OP relying on the GEO database. β-Glycerophosphate disodium salt and L-ascorbic acid were utilized to stimulate differentiation of MC3T3-E1 cells. Cell proliferative, apoptotic abilities, and cell cycle distribution were determined by CCK-8 and flow cytometry experiments. TargetScan and dual-luciferase reporter analysis were employed to predict and verify the targets of miR-23b-3p. Western blot was implemented to detect the expression of CCND1, apoptosis-related proteins, and cell osteogenesis–related proteins. ALP activity of MC3T3-E1 cells was measured using ALP kit. MiR-23b-3p was increased in OP specimens. […]

  • Enhanced KRT13 gene expression bestows radiation resistance in squamous cell carcinoma cells
    on March 1, 2021 at 12:00 am

    Abstract Cancer metastasis and recurrence are potentially lethal. A small number of cancer cell groups called cancer stem cells (CSCs) have both stem cell capacity and cancer-forming ability and are reported to play important roles in cancer metastasis and recurrence. These CSCs are considered to be radiation-resistant (RR). Therefore, understanding the biological effects of radiation on squamous cell carcinoma (SCC) cell lines in vitro and in vivo might be worthwhile to circumvent radiation resistance. Currently, there are no reports on the establishment of RR-SCC cells in serum-free defined culture, which mimics biological mechanisms and prevents instability of using serum in the culture medium. We isolated radiation-resistant strains, designated A431-LDR and A431-HDR, from A431 cells derived from vulval SCC and irradiated them with a total dose of 60 Gy at a low-dose rate (2.2 Gy/d) (RM1000) and a high-dose rate (5 Gy/5.75min) in serum-free defined culture. […]

  • Gga-miR-181a modulates ANP32A expression and inhibits MDCC-MSB-1 cell
    on March 1, 2021 at 12:00 am

    Abstract Marek’s disease (MD), a highly contagious T cell lymphoid neoplasia disease of chickens, causes huge economic losses to the poultry industry. It is the only one tumor disease which can be prevented by vaccine in chickens; therefore, MD is considered to be an excellent model to study the pathogenesis of virus-induced cancer. Recently, abundant evidences have verified that miRNAs are regulators in the process of neoplastic transformation. In our previous study on miRNome analysis of MDV-induced lymphoma in chicken, we found that gga-miR-181a was downregulated drastically in MDV-infected spleens. To further investigate the role of gga-miR-181a in MDV-induced lymphomagenesis, we performed cell migration assay, and the results suggested that gga-miR-181a suppressed the migration of MDV-transformed lymphoid cell (MSB-1). Subsequently, luciferase reporter gene assay revealed that acidic nuclear phosphoprotein 32A (ANP32A) was a functional target gene of […]

  • Advances in 3D neuronal microphysiological systems: towards a functional nervous system on a chip
    on February 1, 2021 at 12:00 am

    Abstract Microphysiological systems (MPS) designed to study the complexities of the peripheral and central nervous systems have made marked improvements over the years and have allowed researchers to assess in two and three dimensions the functional interconnectivity of neuronal tissues. The recent generation of brain organoids has further propelled the field into the nascent recapitulation of structural, functional, and effective connectivities which are found within the native human nervous system. Herein, we will review advances in culture methodologies, focused especially on those of human tissues, which seek to bridge the gap from 2D cultures to hierarchical and defined 3D MPS with the end goal of developing a robust nervous system-on-a-chip platform. These advances have far-reaching implications within basic science, pharmaceutical development, and translational medicine disciplines.

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