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T Titma, R Shimmo, J Siigur, A Kahru
Heavy metals are found naturally on Earth and exposure to them in the living environment is increasing as a consequence of human activity. The toxicity of six different metal oxide nanoparticles (NP) at different points in time was compared using resazurin assay. After incubating Caco2 and A549 cells with 100 μg/mL of Sb2O3, Mn3O4 and TiO2 nanoparticles (NPs) for 24 h no toxic effects were observed while Co3O4 and ZnO NPs had moderate effects and CuO NPs were toxic below 100 μg/mL (24 h EC25 = 11 for A549 and 71 μg/mL for Caco2)...
October 19, 2016: Cytotechnology
Syed Aftab Hussain Shah, Syed Murtaza Hassan Andrabi, Hussain Ahmed, Irfan Zia Qureshi
The objective of the study was to devise a cryoprotection synergism between glycerol and dimethyl sulfoxide (DMSO) for water buffalo spermatozoa. Additionally, the effect of best evolved concentrations of glycerol and DMSO in extender was assessed on in vivo fertility of buffalo spermatozoa. Ejaculates (n = 30) were equally distributed into five aliquots; first aliquot was diluted at 37 °C in extender having 7 % glycerol (control); second aliquot was diluted at 37 °C as well as at 4 °C in extender having 3...
October 19, 2016: Cytotechnology
Ahmad Abu-Khader, Roya Pasha, Gwendoline C D Ward, Gavin Boisjoli, Nicolas Pineault
Engraftment outcomes are strongly correlated with the numbers of hematopoietic stem and progenitor cells (HSPC) infused. Expansion of umbilical cord blood (CB) HSPC has gained much interest lately since infusion of expanded HSPC can accelerate engraftment and improve clinical outcomes. Many novel protocols based on different expansion strategies of HSPC and their downstream derivatives are under development. Herein, we describe the production and properties of serum-free medium (SFM) conditioned with mesenchymal stromal cells derived-osteoblasts (OCM) for the expansion of umbilical CB cells and progenitors...
October 18, 2016: Cytotechnology
Ch Lavanya, M K Sibin, M M Srinivas Bharath, M Jeru Manoj, Manjunatha M Venkataswamy, Dhananjaya I Bhat, K V L Narasinga Rao, G K Chetan
Human telomerase reverse transcriptase (hTERT) gene is a biomarker for the targeted therapy in various cancers. Presence of increased telomerase activity is a common feature of all cancers including glioblastoma. Both RNA and catalytic subunits of hTERT are the target sites for blocking its activity. The current study focuses on the expression of hTERT in glioblastoma and its regulation using two different novel siRNAs (small interfering RNA). Our patient data demonstrated increased expression of hTERT, which could be correlated with carcinogenesis in glioma...
October 18, 2016: Cytotechnology
Yulong Tao, Sang Zhu, Hong Yang, Fei Huang, Hui Fu, Xia Tao
Here, we provide a protocol for reliable isolation and subculture of putative mesenchymal stem cells from mice colons. This method provides a good approach to cultivate and characterize putative colonic mesenchymal stem cells (cMSCs). A high purity of cMSCs can be obtained according to this protocol. The whole isolation processes of cMSCs take about 2 h with two important digestion steps involved. Only with common culture medium, maturation of cMSCs in culture proceeds approximately 2 weeks to allow relevant researches to be conducted...
October 18, 2016: Cytotechnology
İdris Bektaş, Şengül Karaman, Emel Dıraz, Mustafa Çelik
Indigo blue is a natural dye used for thousands of years by civilizations to dye fabric blue and it is naturally obtained from Isatis tinctoria. I. tinctoria is not only used for extraction of indigo blue color but also used medicinally in Traditional Chinese Medicine because of its active compounds. Sodium dithionite (Na2S2O4) is used in dye bath for indigo blue extraction, but this reducing agent and its derivatives are major pollutants of textile industry and subsequently have hazardous influences on public health...
October 18, 2016: Cytotechnology
Serap Sunar, Mustafa Anar, Meryem Sengul, Guleray Agar
In this research, the genotoxic and antigenotoxic effects of methanol extract of Sempervivum armenum (MSA) were studied using micronucleus (MN) test and sister chromatid exchange (SCE) test systems in cultured human peripheral blood cells. According to the SCE and MN tests results, MSA reduced the genotoxic effects of aflatoxin B1. In order to explain the reason for the antigenotoxic effects of MSA, antioxidants levels were determined. Cotreatments of 5, 10, 20 mg/mL concentrations of MSA with aflatoxin B1 decreased the frequencies of SCE, MN and the malondialdehyde level and increased the amount of superoxide dismutase, glutathione and glutathione peroxidase which were decreased by aflatoxin...
October 18, 2016: Cytotechnology
Ulvi K Gursoy, Mervi Gursoy, Eija Könönen, Herman O Sintim, Veli-Jukka Uitto, Stina Syrjänen
In construction of epithelial cells as multilayers, the cells are grown submerged to confluence on fibroblast-embedded collagen gels and, then, lifted to air to promote their stratification. We recently demonstrated that gingival epithelial cells form uniform monolayers on semi-permeable nitrocellulose membranes, supported with a semi-solid growth medium, which allows the cells to grow at an air-liquid-solid interface from the beginning of the culturing protocol. In this study, the aim was to further develop our previous model to form a multilayered gingival epithelial culture model...
October 17, 2016: Cytotechnology
Xinjian Mao, Zhe Chen, Qing Luo, Bingyu Zhang, Guanbin Song
Exposure to microgravity during space flight affects almost all human physiological systems. Migration, proliferation, and differentiation of stem cells are crucial for tissues repair and regeneration. However, the effect of microgravity on the migration potentials of bone marrow mesenchymal stem cells (BMSCs) is unclear, which are important progenitor and supporting cells. Here, we utilized a clinostat to model simulated microgravity (SMG) and found that SMG obviously inhibited migration of rat BMSCs. We detected significant reorganization of F-actin filaments and increased Young's modulus of BMSCs after exposure to SMG...
October 15, 2016: Cytotechnology
Ya-Ching Lin, Guan-Ting Chen, Sheng-Chi Wu
In this study, carbohydrate-chitosan composite including glucose-chitosan, sucrose-chitosan and starch-chitosan with varied carbohydrate concentrations were prepared as carriers for Vero cell culture. Our results show that among these composites, 30 % starch-chitosan composite (STC) were the best carriers for the growth of Vero cells. The initial number of attached cells on the surface of composite carriers did not have any significant effect on subsequent cell production. A higher glucose level in the growth medium during the exponential phase of cell growth, however, played an important factor for cell production...
October 5, 2016: Cytotechnology
Ivette Zegarra Ocampo, Priscila de Queiroz Souza Passos, Luma Ramirez de Carvalho, Camila Ayala Lira da Cruz, Natália Mencacci Esteves-Pedro, Fabiana Medeiros da Silva, Olga Zazuco Higa, Luiz Alberto Pereira Dias, Kayo Okazaki, Daniel Perez Vieira
Micronucleus (MN) assay constitutes a valuable surrogate to the chromosome aberration technique for in vitro testing of the genotoxicity of substances. As test substances, two peptidic compounds (DOTATATE and Ubiquicidin29-41) used in nuclear medicine, were tested for in vitro cytotoxicity and genotoxicity in CHO-K1 cells. None of the compounds showed detectable cytotoxicity (0.5-7.3 ng/mL for DOTATATE and 0.3-4.5 ng/mL for UBI29-41), genotoxicity (0.72, 7.2 and 72.0 ng/ml for DOTATATE and 0.45, 4.5 and 45...
September 29, 2016: Cytotechnology
Yun Gyu Jeong, Jin Sil Lee, Jae Kwon Shim, Won Hur
Multicellular spheroids are obtained in a variety of three-dimensional (3D) culture systems without the use of supporting scaffold. We present here a 3D culture method that resulted in a multicellular sheet under scaffold-free conditions. A floating disk-shaped 3D culture was prepared by magnetic levitation of B16F10 cells that has ingested Fe3O4-containing fibroin microspheres. The melanoma disk grew up to 19 mm in diameter and the thickness was ranged between 80 and 100 μm. The 3D culture was filled with closely packed cells that were proliferating exponentially at a specific growth rate of µ = 0...
September 26, 2016: Cytotechnology
E M Sadeesh, P Sikka, A K Balhara, S Balhara
The study examined the effects of different environmental stress on developmental competence and the relative abundance (RA) of various gene transcripts in oocytes and embryos of buffalo. Oocytes collected during cold period (CP) and hot period (HP) were matured, fertilized and cultured in vitro to blastocyst hatching stage. The mRNA expression patterns of genes implicated in developmental competence (OCT-4, IGF-2R and GDF-9), heat shock (HSP-70.1), oxidative stress (MnSOD), metabolism (GLUT-1), pro-apoptosis (BAX) and anti-apoptosis (BCL-2) were evaluated in immature and matured oocytes as well as in pre-implantation stage embryos...
September 20, 2016: Cytotechnology
Yan-Xin Ren, Jie Yang, Rui-Mei Sun, Li-Juan Zhang, Liu-Fang Zhao, Bao-Zhong Li, Lei Li, Hai-Ting Long, Qiang-Ming Sun, Yun-Chao Huang, Xiao-Jiang Li
The HLA-I antigen processing machinery (APM) plays a crucial role in the anticancer immune response. The loss of surface expression of HLA-I molecules is particularly important as this enables tumor cells to evade recognition and lysis by cytotoxic T-lymphocytes. Transcriptional control of the APM genes is regulated by the nuclear factor kappa B (NF-κB). BCRFl is an Epstein-Barr virus homologue of human IL-10 (hIL-10) and is known as viral IL-10 (vIL-10). vIL-10 shares many immunosuppressive effects with hIL-10 but lacks the immunostimulatory effect of hIL-10...
September 20, 2016: Cytotechnology
Takeru Ota, Mirei Chiba, Haruhide Hayashi
Vibrational stimulation is an accepted non-invasive method used to improve bone remodeling. However, the underlying mechanisms of this phenomenon remain unclear. In this study, we developed a new vibration-loading system to apply vibrational stimulation to cells based on a previously reported in vivo study. We hypothesized that osteoblasts respond to vibrational strain by expressing osteogenic marker genes, such as alkaline-phosphatase (ALP), Runx2, and Osterix. To test our hypothesis, we developed a vibration-loading system to apply a precise vibrational force to an osteoblast culture on a silicone membrane...
September 17, 2016: Cytotechnology
Duyen Thi Do, Nam Nhut Phan, Chih-Yang Wang, Zhengda Sun, Yen-Chang Lin
Indoxyl sulfate (IS) is a digestive intermediate product that is a known indicator of chronic kidney disease. Its toxicity has also been suggested to accelerate chronic kidney disease. Recently, mesenchymal stem cells (MSCs) have been confirmed as a potential treatment in kidney regeneration. To determine the universal alteration in gene expression, we combined high-throughput microarray technology and in vitro culture of adipose-derived mesenchymal stem cells at different doses of IS (20, 40, 60 mg/l). We found that indoxyl sulfate has a remarkable interconnection with stem cell and calcium/calmodulin-dependent kinase pathways...
August 22, 2016: Cytotechnology
Yasuhiro Takagi, Tomomi Yamazaki, Kenji Masuda, Shigeaki Nishii, Bunsei Kawakami, Takeshi Omasa
Chinese hamster ovary (CHO) cell lines are widely used for therapeutic protein production. When a transgene is integrated into the genome of a CHO cell, the expression level is highly dependent on the site of integration because of positional effects such as gene silencing. To overcome negative positional effects and establish stable CHO cell lines with high productivity, several regulatory DNA elements are used in vector construction. Previously, we established the CHO DR1000L-4N cell line, a stable and high copy number Dhfr gene-amplified cell line...
August 20, 2016: Cytotechnology
Shin-Ichi Adachi, Fumiaki Yoshizawa, Kazumi Yagasaki
Hyperuricemia is characterized by the high uric acid (UA) level in serum (or plasma) and has been considered to be an important risk factor for gout. In the present study, we have attempted to construct an assay system for UA production in vitro employing cultured AML12 hepatocytes. UA levels in balanced salt solution (BSS) in the presence of UA precursor nucleosides, adenosine, inosine, guanosine and xanthine, at 12.5, 25, and 100 µM were significantly higher than BSS alone and their effects were dose-dependent, while all the UA precursors did not significantly increase intracellular UA levels...
August 12, 2016: Cytotechnology
Ebru Hacıosmanoğlu, Başak Varol, Bilge Özerman Edis, Muhammet Bektaş
Eukaryotic elongation factor 2 (eEF2) plays an important role in eukaryotic polypeptide chain elongation. Adenosine diphosphate (ADP)-ribosylation is a post-translational modification reaction that catalyzes the transfer of ADP-ribose group to eEF2 and this causes the inhibition of protein synthesis. Indeed, in the absence of diptheria toxin, endogenous ADP-ribosylation can occur. eEF2 is phosphorylated by eEF2 kinase which prevents binding to ribosomes thus inhibiting its activity. Increase in endogenous ADP-ribosylation level approximately 70-75 % was observed in IL-1β treated HUVECs...
August 10, 2016: Cytotechnology
Xudong Ao, Rula Sa, Jie Wang, Rinuo Dao, Huimin Wang, Haiquan Yu
DNA methylation in mammals is an epigenetic marker and necessary for normal embryogenesis. The global genomic demethylation of 5-methylcytosine occurs during the first cell cycle following fertilization. Activation-induced cytidine deaminase (AID), which is well-known for the function in antibody diversification, has been implicated to play a role in active demethylation, but its role in cell reprogramming and its crosstalk with other DNA demethylation mechanism need to be clarified. In this study, the dynamic epigenetic regulation of cell pluripotency and embryo development by AID in bovine preimplantation embryos was investigated...
August 9, 2016: Cytotechnology
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