Recent Articles

3D Cell CultureBelow 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.


  • Influence of standard culture conditions and effect of oleoresin from the microalga Haematococcus pluvialis on splenic cells from healthy Balb/c mice — a pilot study
    on December 7, 2023 at 12:00 am

    Abstract In this work, we used splenocytes from healthy mice to study the effects of the two most commonly used cell culture media (A, B) with different compositions of redox reagents. The incubation of cells for 24 h resulted in a significant decrease in viability and metabolic activity of splenocytes, and the negative effects of incubation in medium B were more pronounced. In standard conditions, oxidative stress in cells was manifested by reduced mitochondrial potential, and this effect correlated with the transition of 58.3% of cells to the early stage of apoptosis under reducing conditions of medium A and up to 66.1% of cells under super-reducing conditions in medium B, suggesting altered cell physiology. High levels of ROS/RNS activated transcription factor Nrf2, superoxide dismutase 1, and catalase. The higher mRNA levels of these genes were under the conditions of medium B, whose super-reducing environment in combination with the environment of conventional […]

  • Downregulation of BUBR1 regulates the proliferation and cell cycle of breast cancer cells and increases the sensitivity of cells to cisplatin
    on December 4, 2023 at 12:00 am

    Abstract Breast cancer (BC) is a significant tissue for women’s health worldwide. The spindle assembly checkpoint protein family includes BUBR1 (Bub1-related kinase or MAD3/Bub1b). High expression of BUBR1 promotes cell cycle disorders, leading to cell carcinogenesis and cancer progression. However, the underlying molecular mechanism and the role of BUBR1 in BC progression are unclear. The published dataset was analyzed to evaluate the clinical relevance of BUBR1. BUBR1 was knocked down in BC cells using shRNA. The CCK-8 assay was used to measure the cell viability, and mRNA and protein expression levels were detected by RT-qPCR and Western blot (WB). Cell apoptosis and cell cycle were detected by flow cytometry. Subcutaneous xenograft model was used to assess in vivo tumor growth. BUBR1 was found to be highly expressed in BC. The high expression of BUBR1 was associated with poor prognosis of BC patients. Upon BUBR1 knockdown using shRNA, the proliferation and […]

  • Epigallocatechin gallate inhibits ovarian cancer cell growth and induces cell apoptosis via activation of FOXO3A
    on December 1, 2023 at 12:00 am

    Abstract Epigallocatechin gallate (EGCG), a bioactive component in tea, displays broad anti-cancer effects. Our study was designed to evaluate the anti-cancer effects of EGCG on ovarian cancer and explored the underlying molecular mechanisms. To evaluate the in vitro inhibitory effects of EGCG against ovarian cancer, MTT assay, colony formation assay, apoptosis assay, and wound healing assay, were performed. Besides, the inhibitory effects of EGCG on tumor growth in the xenograft animal model were evaluated by measuring tumor volume and tumor weight. Moreover, Western blotting and qPCR were used to evaluate the levels of target genes and proteins. Treatment with EGCG inhibited cell migration and cell survival, and promoted cell apoptosis in A2780 and SKOV3 cells. Interestingly, treatment with EGCG inhibited the tumor growth in the xenograft animal model. The mechanistic study revealed that treatment with EGCG induced the activation of FOXO3A and suppressed the […]

  • Myricetin ameliorates the IL-21-induced tumorigenic phenotype of adjuvant-induced arthritis FLS by modulating the choline kinase signaling cascade
    on November 30, 2023 at 12:00 am

    Abstract The synovial intimal lining is mainly governed by fibroblast-like synoviocytes (FLS), which portray a transformed tumor-like phenotype in rheumatoid arthritis (RA). Among the diverse cytokines that engender FLS, interleukin-21 (IL-21) was reported to stimulate hyperproliferation and perpetuate inflammation. Recently, choline kinase (ChoKα) has been reported to be an essential enzyme aiding RA-FLS hyperproliferation by altering phosphatidylcholine biosynthesis. The current study aimed to elucidate the therapeutic efficacy of myricetin, a flavonoid, in abating the IL-21-induced tumor-like phenotype of adjuvant-induced arthritis (AIA)-FLS via the ChoKα signaling cascade. Our results showed that myricetin suppressed IL-21 receptor expression and activation of the ChoKα signaling cascade (N-Ras, Ral-GDS, and PI3K) in IL-21-induced AIA-FLS. Consequently, myricetin treatment decreased ChoKα and PLD2 enzymatic activity and inhibited the proliferative, migratory, […]

  • Correction: CD142 promotes trophoblast cell migration by inhibiting BCL2-related autophagic degradation of IL-8
    on November 29, 2023 at 12:00 am
  • Cytotoxicity of the field lampricide 3-trifluoromethyl-4-nitrophenol (TFM) in tadpole cell lines from North American frogs
    on November 27, 2023 at 12:00 am

    Abstract The common field lampricide, 3-trifluoromethyl-4-nitrophenol (TFM), is used to treat streams and creeks infested with highly invasive and destructive sea lamprey (Petromyzon marinus) in the tributaries of the Great Lakes. Unfortunately, amphibian deaths have been reported following stream treatments with TFM. Larval amphibians (tadpoles) are more susceptible to TFM toxicity than adult amphibians. The aim of this study was to test the toxicity of TFM in eight new tadpole cell lines from the green frog (Lithobates clamitans), wood frog (Lithobates sylvaticus), and American toad (Anaxyrus americanus). A cell viability bioassay using two fluorescent dyes, Alamar Blue and CFDA-AM, was performed following 24-h and 72-h exposures to a range of TFM concentrations. In general, TFM exposure reduced Alamar Blue fluorescence more rapidly than CFDA-AM fluorescence in tadpole cells, suggesting that Alamar Blue is perhaps a better diagnostic indicator of cell health for […]

  • Establishment of SLC7A11‐knockout mouse and its preliminary investigation in melanoma
    on October 1, 2023 at 12:00 am

    Abstract Solute carrier family 7 member 11 (SLC7A11)/xCT is an amino acid transporter that mediates the cystine uptake and glutamate export, participates in several malignant tumors’ progression. However, the role of SLC7A11 on the occurrence and development of melanoma still remains unclear. Here, the transcribed mRNA encoding for Cas9 and sgRNA targeting SLC7A11 in vitro were microinjected into zygotes, to establish the SLC7A11 knockout (KO) mice (SLC7A11−/−). Further, we conducted melanoma-bearing mice using the metastatic melanoma cell line (B16-F10) to observe the melanoma development. There was no off-target in KO mice detected by T7E1 cleavage assay. The results showed that the tumor volume of KO mice was significantly lower than that of SLC7A11+/+ (WT) mice at 8d, 10d, 12d, 14d, and 16d (P < 0.05). The tumors of WT appeared to more disorganized morphology, more unbalanced nuclear-cytoplasmic ratio, less defined boundary, and increased tumor […]

  • Ulinastatin ameliorates podocyte ferroptosis via regulating miR-144-3p/SLC7A11 axis in acute kidney injury
    on October 1, 2023 at 12:00 am

    Abstract Ferroptosis is a newly discovered form of cell death characterized by intracellular iron accumulation and subsequent lipid peroxidation, which has been identified in various pathological processes, such as acute kidney injury (AKI). Ulinastatin (UTI), known as an antioxidant and anti-inflammatory, has been reported to prevent kidney injury. Here, we investigated the protective effects of UTI on LPS-induced podocyte ferroptosis in vivo and in vitro. Conditionally immortalized mouse podocyte was exposed to LPS in the presence or absence of UTI in vitro for 48 h. The levels of reactive oxygen species (ROS) and intracellular Fe2+ were detected to value the effect of UTI treatment on the podocyte cell ferroptosis. We also evaluated the influence of UTI on kidney injury in vivo. LPS-induced mice were treated with vehicle or UTI at 50 U/g/d for 6 wk. We identified the important function of UTI in repressing ferroptosis and ameliorating podocyte injury. The […]

  • Anti-oxidative activities and anti-ferroptosis of conditioned medium from umbilical cord mesenchymal stem cells
    on October 1, 2023 at 12:00 am

    Abstract The therapeutic effect of MSC is closely related to its antioxidant capacity. There is no uniform standard for evaluating the antioxidant capacity of MSC. In this study, we compared the antioxidant capacity of control medium (CON) and conditioned medium (CM) from umbilical cord mesenchymal stem cells cultured for 48 h, about total antioxidant capacity, DPPH scavenging capacity, O2- and hydroxyl radical inhibiting capacity, and the detection of antioxidant enzymes including superoxide dismutase, glutathione peroxidase, and catalase, and resistance to cellular oxidative damage caused by H2O2, SNAP, erastin, and RSL3. The results showed that CM had better DPPH scavenging capacity than CON. No significant differences were observed in antioxidant enzymes. CM did not resist the oxidative damage induced by H2O2 and SNAP, but it had a strong resistance to ferroptosis induced by erastin and RSL3, indicating that CM had excellent resistance to cell lipid […]

  • Ghrelin inhibits NLRP3 inflammasome activation by upregulating autophagy to improve Alzheimer’s disease
    on October 1, 2023 at 12:00 am

    Abstract Nod-like receptor protein 3 (NLRP3) inflammasome, autophagy, and the aggregation of β-amyloid (Aβ) are key factors in Alzheimer’s disease (AD) development. Ghrelin has shown promise in providing neuroprotection for AD. However, the mechanism underlying ghrelin’s ability to improve AD by modulating autophagy and the NLRP3 inflammasome requires further clarification. Primary hippocampus neurons and mice were stimulated with Aβ1-42 to create an in vitro and in vivo AD model, followed by ghrelin administration for intervention. Additionally, we subjected the cells to 3-methyladenine (3-MA) treatment. Neuron morphology, microtubule-associated protein 2 (MAP-2) expression, apoptosis, cytokine levels, and protein expression were measured using various techniques. The escape latency of mice was assessed using the Morris water maze (MWM) test, and histopathology of the hippocampus was determined using hematoxylin–eosin staining. At 1–100 nM, ghrelin […]

  • Curculigoside attenuates osteoporosis through regulating DNMT1 mediated osteoblast activity
    on October 1, 2023 at 12:00 am

    Abstract This work aims to study the function of curculigoside in osteoporosis and explore whether DNMT1 is closely involved in osteoblast activity. After OB-6 osteoblasts were treated with hydrogen peroxide (H2O2), a curculigoside treatment group was set up and a series of biological tests including MTT, flow cytometry, western blotting, ROS fluorescence intensity, mitochondrial membrane potential, and ELISA experiments were performed to verify the effect of curculigoside on the activity of osteoblasts. Then, alkaline phosphatase (ALP) activity, alizarin red staining, PCR, and western blotting assays were performed to detect the effects of curculigoside on osteoblast function. By constructing DNMT1 knockdown and overexpression OB-6 cell lines, the effect of DNMT1 on osteoblast function was verified. In addition, the expression level of Nrf2 in each group was detected to speculate the mechanism of DNMT1 in osteoporosis. The cell activity and level of bcl-2 and SOD […]

  • Drynaria Naringin alleviated mechanical stress deficiency-caused bone loss deterioration via Rspo1/Lgr4-mediated Wnt/β-catenin signalling pathway
    on October 1, 2023 at 12:00 am

    Abstract  Osteoporosis is a metabolic condition distinguished by the degradation of bone microstructure and mechanical characteristics. Traditional Chinese medicine (TCM) has been employed in China for the treatment of various illnesses. Naringin, an ingredient found in Drynariae TCM, is known to have a significant impact on bone metabolism. For this research, we studied the precise potential effect of Drynaria Naringin on protecting against bone loss caused by stress deficiency. In this study, a tail-suspension (TS) test was performed to establish a mouse model with hind leg bone loss. Some mice received subcutaneous injections of Drynaria Naringin for 30 d. Trabecular bone microarchitecture was evaluated using micro-computed tomography analysis and bone histological analysis. Bone formation and resorption markers were quantified in blood samples from mice or in the supernatant of MC3T3-E1 cells by ELISA analysis, Western blotting, and PCR. Immunofluorescence was […]

  • Ecliptasaponin A attenuates renal fibrosis by regulating the extracellular matrix of renal tubular cells
    on October 1, 2023 at 12:00 am

    Abstract Renal fibrosis is the most common manifestation of end-stage renal disease (ESRD), including diabetic kidney disease (DKD), but there is no effective treatment in renal fibrosis. Natural products are a rich source of clinical drug research and have been used in the clinical research of various diseases. In this study, we searched for traditional Chinese medicine monomers that attenuate fibrosis and assessed their effect on the fibrosis marker connective tissue growth factor (CTGF) in cells which we found ecliptasaponin A. Subsequently, we evaluated the effect of ecliptasaponin A on renal fibrosis in the classic renal fibrosis unilateral ureteral obstruction (UUO) mouse model and found that ecliptasaponin A could reduce the renal collagen fiber deposition and renal extracellular matrix (ECM) protein expression in UUO mice. In vitro, ecliptasaponin A can inhibit ECM protein expression in human kidney-2 (HK-2) cells induced by transforming growth […]

  • Identification of highly active compounds from insecticidal plant Oroxylum indicum L. (Vent.) and the induction of apoptosis by lapachol on Sf9 cells
    on October 1, 2023 at 12:00 am

    Abstract The extraction of biopesticides from plants has become a promising field for agricultural development. To explore a high-efficiency and viable method for the screening of plant compounds with insecticidal activity, we screened for active ingredients in the insecticidal plant, Oroxylum indicum L. Vent, using Sf9 cells. A CCK-8 cytotoxicity assay kit was used for high-throughput screening of 34 compounds contained in O. indicum. The apoptosis-inducing effect of the highly cytotoxic compound on Sf9 cells was investigated by morphological characterization using inverted microscopy, caspase-3 activity assay, and DNA gel electrophoresis. Finally, the biological activity of compounds against aphids was evaluated using the leaf-pest dipping methods and leaf dipping methods. Results showed that among the main compounds identified, lapachol, chrysin, and baicalein had good proliferation inhibitory effects on Sf9 cells, with their recorded IC50 being 11.53 mg/L, […]

  • Vardenafil alleviates cigarette smoke-induced chronic obstructive pulmonary disease by activating autophagy via the AMPK/mTOR signalling pathway: an in vitro and in vivo study
    on October 1, 2023 at 12:00 am

    Abstract Chronic obstructive pulmonary disease (COPD) has always attracted global attention with its high prevalence, incidence rate, and mortality. Exposure to cigarette smoke is one of main causes of COPD. Therefore, it is still necessary to study its pathogenesis and find new therapeutic strategies for early COPD prevention and treatment. Vardenafil, a type 5 phosphodiesterase (PDE5) inhibitor, is known to have an efficient therapy in some cardiovascular, pulmonary, and vascular diseases, which is an important mechanism for COPD. However, it still loss relevant research on whether vardenafil is effective in COPD and its mechanism. In this study, the cigarette smoke inhalation was performed to establish cigarette smoke-induced COPD model using C57BL/6 mice and 16HBE cells were treated with cigarette smoke extract (CSE). Mice were treated with vardenafil for 30 d. Then condition of lung injury was evaluated using histological analysis. The content of cytokines and […]

  • Punicalagin relieves lipotoxic injuries on pancreatic β-cells via regulating the oxidative stress and endoplasmic reticulum stress-mediated apoptosis
    on September 1, 2023 at 12:00 am

    Abstract Cellular toxicity of hyperlipidemia has been long considered a major cause of various intractable disease such as diabetes. Discovering lipotoxicity antagonist with high efficiency and low side effects is of importance to develop therapeutics for relevant diseases. In the current study, we evaluate the anti-lipotoxic potential of punicalagin (PU) on pancreatic cells and investigate its underpinning mechanism involved. The administration of PU effectively improved cell viability, quenched intracellular reactive oxygen species, alleviated lipid peroxidation, and enhanced cellular antioxidative capacity in RINm5F cells stimulated by sodium palmitate. Besides that, PU treatment significantly inhibited the overload of mitochondrial calcium ions; alleviated the activation of endoplasmic reticulum (ER) stress mediators including glucose-regulated protein 78, protein kinase RNA-like ER kinase, eukaryotic initiation factor 2α, activating transcription factor 6, […]

  • Platelet factor 4 (PF4) induces cluster of differentiation 40 (CD40) expression in human aortic endothelial cells (HAECs) through the SIRT1/NF-κB/p65 signaling pathway
    on September 1, 2023 at 12:00 am

    Abstract PF4 is a pro-atherosclerotic molecule. Endothelial CD40, upon binding to its ligand CD40L, induces endothelial cell (EC) activation, which is a vital pathophysiological process in the initiation and progression of atherosclerosis. However, the relationship between PF4 and endothelial CD40 remains elusive. This study aims to investigate whether and how PF4 affects endothelial CD40 expression using primary HAECs. PF4 treatment down-regulated sirtuin 1 (SIRT1) expression but upregulated the expression of acetylated NF-κB p65 (Ac-p65) and CD40 in HAECs in a concentration- and time-dependent manner. Pretreatment with SIRT1 agonist (SRT1720 or RSV) or SIRT1-overexpressing lentivirus attenuated PF4-induced Ac-p65 and CD40 expression in HAECs, whereas preincubation with SIRT1 antagonist (NAM or EX527) or SIRT1 shRNA had the opposite effect. To investigate whether NF-κB/p65 signaling pathway modulates CD40 expression in PF4-treated HAECs, PDTC, a NF-κB […]

  • HIF-1α-induced upregulation of m6A reader IGF2BP1 facilitates peripheral nerve injury recovery by enhancing SLC7A11 mRNA stabilization
    on September 1, 2023 at 12:00 am

    Abstract The recovery of peripheral nerve injury (PNI) is not ideal in clinic. Our previous study revealed that hypoxia treatment promoted PNI repair by inhibiting ferroptosis. The aim of this study was to investigate the underlying molecular mechanism of HIF-1α in hypoxia-PNI recovery. M6A dot blot was used to determine the total level of m6A modification. Besides, HIF-1α small interfering RNA (siRNA) or IGF2BP1 overexpression vector was transfected into dorsal root ganglion (DRG) neurons to alter the expression of HIF-1α and IGF2BP1. Subsequently, MeRIP-PCR analysis was applied to validate the m6A methylation level of SLC7A11. We demonstrated the hypoxia stimulated HIF-1α-dependent expression of IGF2BP1 and promoted the overall m6A methylation levels of DRG neurons. Overexpression of HIF-1α increased the expressions of neurotrophic factors including nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), and glial-derived neurotrophic factor […]

  • In vitro inhibitory effect of zingerone on TNFα-stimulated fibroblast-like synoviocytes
    on September 1, 2023 at 12:00 am

    Abstract Targeting Fibroblast-like synoviocytes (FLSs) is an attractive complementary approach for RA therapy. This study aimed to investigate the inhibitory effects of zingerone on TNFα-induced arthritic FLSs. MTS, EdU, wound healing, DHE staining and real-time PCR were used to determine the effects of zingerone on the destructive behaviors of arthritic FLSs induced by TNFα. Western blot analysis was used to analyze cell signaling pathways. Zingerone treatment significantly inhibited TNFα-induced proliferation, migration, ROS formation and pro-inflammatory cytokines expression of FLSs. Molecular mechanism studies revealed that zingerone could suppress TNFα-induced activations of MAPKs (ERK, JNK and p38) in arthritic FLSs. Zingerone attenuated pathological features of FLSs via MAPKs pathways, indicating its potential as a complementary or alternative drug for RA therapy.

  • Culture methods focusing on bile canalicular formation using primary human hepatocytes in a short time
    on September 1, 2023 at 12:00 am

    Abstract The development of models for predicting hepatotoxicity is warranted, as the hepatotoxicity risk of 38–51% of compounds is undetectable in nonclinical studies. Cholestatic drug-induced liver injury (DILI) is a condition in which bile acids are abnormally excreted into the capillary bile canaliculi and are accumulated in hepatocytes, caused by the inhibition of bile salt export pump (BSEP), a transporter that is mainly associated with excretion of bile acids. Although laboratory animals are used as models, the use of human-derived cells is required owing to species differences. Unfortunately, primary human hepatocytes (PHHs) show rapid loss of function in culture and difficulties in forming bile canaliculi. Therefore, we aimed to develop an in vitro culture method for the efficient formation of bile canaliculi and for assessing the function of BSEP in PHHs. Here, PHHs were cultured from 1 h after thawing to day 2 with Z-VAD-FMK, a total caspase […]

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