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Cytotechnology

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https://www.readbyqxmd.com/read/28799096/a-quantitative-and-qualitative-rna-expression-profiling-assay-for-cell-culture-with-single-cell-resolution
#1
Petra Kraus, Rachel Yerden, Darren Sipes, Shantanu Sur, Thomas Lufkin
Cells are often characterized by their gene expression profile. However, commonly used methods to detect mRNA require cell pooling and could therefore mask differences in gene expression within heterogeneous cell populations. q(2)PISH allows for the analysis of both qualitative and quantitative (q(2)) gene expression on cultured cells for quality control measures with single cell resolution. q(2)PISH was optimized for the subsequent use of two alkaline phosphatase substrates in combination with a cell nucleus count to allow for accurate quantification of gene expression per cell and simultaneously qualitative assessment of potential culture population drift or heterogeneity...
August 10, 2017: Cytotechnology
https://www.readbyqxmd.com/read/28780625/markers-associated-with-neuron-specific-ube3a-imprinting-during-neuronal-differentiation-of-mouse-embryonic-stem-cells
#2
Masamitsu Eitoku, Hidemasa Kato, Narufumi Suganuma, Hidenori Kiyosawa
Understanding gene expression in the brain requires allele-specific transcriptome analysis because of the presence of neuron-specific imprinted genes, which are expressed in a neuron-specific and parent-of-origin-specific manner. Ube3a is a neuron-specific imprinted gene with an expression pattern that changes from biallelic to maternal only (Ube3a imprinting) during differentiation. Because Ube3a imprinting occurs only in neurons, it has the potential to be a marker to assess the quality of neurons produced by in vitro neuronal differentiation of embryonic stem cells (ESCs)...
August 5, 2017: Cytotechnology
https://www.readbyqxmd.com/read/28779292/the-use-of-an-artificial-neural-network-to-model-the-infection-strategy-for-baculovirus-production-in-suspended-insect-cell-cultures
#3
Antonio Contreras-Gómez, Alba Beas-Catena, Asterio Sánchez-Mirón, Francisco García-Camacho, Emilio Molina Grima
Since the infection strategy in the baculovirus-insect cell system mostly affects production of the vector itself or the target product, and given that individual infection parameters interact with each other, the optimal combination must be established for each such specific system. In this work an artificial neural network was used to model infection strategy, including the cell concentration at infection, the multiplicity of infection, the medium recycle, and agitation intensity, and to evaluate the relative importance of each factor in the baculovirus production obtained...
August 4, 2017: Cytotechnology
https://www.readbyqxmd.com/read/28756527/synergistic-tumor-suppression-by-a-perilla-frutescens-derived-methoxyflavanone-and-anti-cancer-tyrosine-kinase-inhibitors-in-a549-human-lung-adenocarcinoma
#4
Amer Ali Abd El-Hafeez, Takashi Fujimura, Rikiya Kamei, Noriko Hirakawa, Kenji Baba, Kazuhisa Ono, Seiji Kawamoto
Anti-cancer tyrosine kinase inhibitors (TKIs) are effective in many types of cancers including non-small cell lung cancer, while appearance of TKI-resistant tumors suggests a need for the development of their potentiation strategies. We have previously shown that a methoxyflavanone derivative from the Asian medicinal herb Perilla frutescens (Perilla-derived methoxyflavanone; PDMF) shows a prominent anti-tumor activity against A549 human lung adenocarcinoma. Here we show that PDMF and anti-cancer TKIs (nilotinib, bosutinib, dasatinib, and ponatinib) synergistically suppress proliferation of A549 cells...
July 29, 2017: Cytotechnology
https://www.readbyqxmd.com/read/28755030/the-human-vascular-endothelial-cell-line-huv-ec-c-harbors-the-integrated-hhv-6b-genome-which-remains-stable-in-long-term-culture
#5
Setsuko Shioda, Fumio Kasai, Midori Ozawa, Noriko Hirayama, Motonobu Satoh, Yousuke Kameoka, Ken Watanabe, Norio Shimizu, Huamin Tang, Yasuko Mori, Arihiro Kohara
Human herpes virus 6 (HHV-6) is a common human pathogen that is most often detected in hematopoietic cells. Although human cells harboring chromosomally integrated HHV-6 can be generated in vitro, the availability of such cell lines originating from in vivo tissues is limited. In this study, chromosomally integrated HHV-6B has been identified in a human vascular endothelial cell line, HUV-EC-C (IFO50271), derived from normal umbilical cord tissue. Sequence analysis revealed that the viral genome was similar to the HHV-6B HST strain...
July 28, 2017: Cytotechnology
https://www.readbyqxmd.com/read/28752496/investigations-on-in-vitro-anti-carcinogenic-potential-of-l-carnosine-in-liver-cancer-cells
#6
Minghui Ding, Guihua Jiao, Haizhou Shi, Yanrong Chen
This study was carried out to investigate the anti-carcinogenic effect of L-carnosine in human carcinoma cells (SNU-423). The SNU-423 cancer cells were cultured at a density of 2 × 10(4) cells/well in Dulbecco modified Eagle medium. After 24 h of adherence, the cells were treated with L-carnosine (0.2 and 1 mg/mL) for 48 h. Then, cell viability was assessed by sulforhodamine assay, while mitochondrial dysfunction was measured by fluorescence microscopy using chromatin-specific dye Hoechst 33258. Intracellular levels of ROS were assayed by fluorescence spectroscopy with 2',7'-dichlorofluorescein diacetate (DCFDA)...
July 27, 2017: Cytotechnology
https://www.readbyqxmd.com/read/28710570/a-methoxyflavanone-derivative-from-the-asian-medicinal-herb-perilla-frutescens-induces-p53-mediated-g2-m-cell-cycle-arrest-and-apoptosis-in-a549-human-lung-adenocarcinoma
#7
Amer Ali Abd El-Hafeez, Takashi Fujimura, Rikiya Kamei, Noriko Hirakawa, Kenji Baba, Kazuhisa Ono, Seiji Kawamoto
Perilla frutescens is an Asian dietary herb consumed as an essential seasoning in Japanese cuisine as well as used for a Chinese medicine. Here, we report that a newly found methoxyflavanone derivative from P. frutescens (Perilla-derived methoxyflavanone, PDMF; 8-hydroxy-5,7-dimethoxyflavanone) shows carcinostatic activity on human lung adenocarcinoma, A549. We found that treatment with PDMF significantly inhibited cell proliferation and decreased viability through induction of G2/M cell cycle arrest and apoptosis...
July 14, 2017: Cytotechnology
https://www.readbyqxmd.com/read/28702859/spirulina-platensis-prevents-high-glucose-induced-oxidative-stress-mitochondrial-damage-mediated-apoptosis-in-cardiomyoblasts
#8
Pratiksha Jadaun, Dhananjay Yadav, Prakash Singh Bisen
The current study was undertaken to study the effect of Spirulina platensis (Spirulina) extract on enhanced oxidative stress during high glucose induced cell death in H9c2 cells. H9c2 cultured under high glucose (33 mM) conditions resulted in a noteworthy increase in oxidative stress (free radical species) accompanied by loss of mitochondrial membrane potential, release of cytochrome c, increase in caspase activity and pro-apoptotic protein (Bax). Spirulina extract (1 μg/mL), considerably inhibited increased ROS and RNS levels, reduction in cytochrome c release, raise in mitochondrial membrane potential, decreased the over expression of proapoptotic protein Bax and suppressed the Bax/Bcl2 ratio with induced apoptosis without affecting cell viability...
July 12, 2017: Cytotechnology
https://www.readbyqxmd.com/read/28689280/analysis-of-toxicity-effects-of-di-2-ethylhexyl-phthalate-exposure-on-human-bronchial-epithelial-16hbe-cells
#9
Yue Ma, Yinsheng Guo, Shuang Wu, Ziquan Lv, Qian Zhang, Xing Xie, Yuebin Ke
Recent studies have indicated that Di-(2-ethylhexyl) phthalate (DEHP), the most commonly used plasticizer in daily-life products, could be dispersed in indoor air and induce human exposure via inhalation. DEHP has been reported to have effects on the respiratory system in both animal and human researches. The toxicity effects of DEHP exposure on cell proliferation, cell cycle progression, apoptosis, global DNA methylation and the expression levels of DNA methyltransferases (DNMTs) were investigated in this study, using human epithelial cell line 16HBE as an in vitro model...
July 8, 2017: Cytotechnology
https://www.readbyqxmd.com/read/28689279/three-dimensional-culture-of-a-genetically-modified-hepatoma-cell-line-using-macroporous-gelatin-beads
#10
Jane Marie Tonello, Saori Kawashima, Kazuki Sato, Yoshinori Kawabe, Akira Ito, Masamichi Kamihira
Hepatoma cells are a candidate cell source for bio-artificial livers. However, they exhibit reduced liver functions compared with primary hepatocytes. In our previous study, genetically engineered mouse hepatoma cells were created by transduction with vectors mediating inducible overexpression of eight liver-enriched transcription factors. Upon the induction of the liver-enriched transcription factors transduced, the cells expressed both phenotypic and genotypic liver functions at high levels. In the present study, we performed three-dimensional culture of these cells using macroporous gelatin beads...
July 8, 2017: Cytotechnology
https://www.readbyqxmd.com/read/28625011/liver-epithelioid-progenitor-cells-derived-from-fetal-luxi-bovine-alleviate-liver-fibrosis
#11
Kunfu Wang, Hao Liu, Jinjuan Yang, Caiyun Ma, Zebiao Zhang, Dong Zheng, Weijun Guan
Liver epithelioid progenitor cells (LEPCs) have important roles in liver therapy because of their hepatic differentiation potency in vitro and in vivo. Despite many researches on humans, mice, and rats, equivalent progenitor cells derived from bovine are relatively rare. The purpose of our current study is to characterize bovine LEPCs, and research on the cure potency of this heteroplastic progenitor cells on mice liver fibrosis. We have used collagenase IV digesting and differential adhesion method to isolate slabstone shape, EpCAM, LGR5, NCAM1 and SOX9 positive progenitor cells from fetal Luxi bovine liver...
June 17, 2017: Cytotechnology
https://www.readbyqxmd.com/read/28608258/extracts-of-chrysanthemum-zawadskii-attenuate-oxidative-damage-to-vascular-endothelial-cells-caused-by-a-highly-reducing-sugar
#12
Hyun-Sook Kim
Endothelial cells are considered candidates for involvement in the pathogenesis of diabetic vascular complications, and prevention of endothelial cell damage may be important in pharmacological attempts to prevent such complications. In the present study, I explored whether extracts of Chrysanthemum zawadskii (CZE) could prevent oxidative damage and dysfunction of a vascular endothelial cell line caused by the highly reducing sugar, 2-deoxy-D-ribose (dRib), and dysfunction of a vascular endothelial cell line...
June 12, 2017: Cytotechnology
https://www.readbyqxmd.com/read/28608257/mutagenic-and-genotoxic-effects-of-anilofos-with-micronucleus-chromosome-aberrations-sister-chromatid-exchanges-and-ames-test
#13
Dilek Akyil, Muhsin Konuk, Yasin Eren, Recep Liman, Esra Sağlam
We aimed to evaluate the mutagenic effect of Anilofos, organophosphate pesticide, by using Ames/Salmonella/microsome test. Its cytotoxic and genotoxic effects were also determined by chromosome aberration (CA), sister chromatid exchange (SCE) and micronucleus (MN) test in human peripheral blood lymphocytes. In the Ames test, five different concentrations of Anilofos were examined on TA97, TA98, TA100 and TA102 strains in the absence and presence of S9 fraction. According to the results all concentrations of this pesticide have not shown any mutagenic activity on TA97, TA100 and TA102 strains in the absence and presence of S9 fraction...
June 12, 2017: Cytotechnology
https://www.readbyqxmd.com/read/28601931/cardiotrophin-1-stimulates-the-neural-differentiation-of-human-umbilical-cord-blood-derived-mesenchymal-stem-cells-and-survival-of-differentiated-cells-through-pi3k-akt-dependent-signaling-pathways
#14
Longying Peng, Xiaomei Shu, Changhui Lang, Xiaohua Yu
Cardiotrophin-1 (CT1) plays an important role in the differentiation, development, and survival of neural stem cells. In this study, we analyzed its effects on the stimulation of human umbilical cord blood-derived mesenchymal stem cells in terms of their potential to differentiate into neuron-like cells, their survival characteristics, and the molecular mechanisms involved. The treatment of cells with neural induction medium (NIM) and CT1 generated more cells that were neuron-like and produced stronger expression of neural-lineage markers than cells treated with NIM and without CT1...
June 10, 2017: Cytotechnology
https://www.readbyqxmd.com/read/28584932/modifications-of-a-signal-sequence-for-antibody-secretion-from-insect-cells
#15
Yuki Ohmuro-Matsuyama, Hideki Yamaji
Monoclonal antibodies and antibody fragments have recently been developed for use in diverse diagnostic and therapeutic applications. Insect cells can efficiently secrete recombinant proteins such as antibody molecules through post-translational processing and modifications that are similar to those performed in mammalian cells. In eukaryotic cells, the signal sequence in a nascent polypeptide is recognized by the signal recognition particle, and the polypeptide is then folded and modified in the endoplasmic reticulum...
June 5, 2017: Cytotechnology
https://www.readbyqxmd.com/read/28573544/lipocalin-2-enhances-mesenchymal-stem-cell-based-cell-therapy-in-acute-kidney-injury-rat-model
#16
Mehryar Habibi Roudkenar, Raheleh Halabian, Hossein Abdul Tehrani, Fatemeh Amiri, Ali Jahanian-Najafabadi, Amaneh Mohammadi Roushandeh, Zahra Abbasi-Malati, Yoshikazu Kuwahara
Acute kidney injury (AKI) is one of the most common health-threatening diseases in the world. There is still no effective medical treatment for AKI. Recently, Mesenchymal stem cell (MSC)-based therapy has been proposed for treatment of AKI. However, the microenvironment of damaged kidney tissue is not favorable for survival of MSCs which would be used for therapeutic intervention. In this study, we genetically manipulated MSCs to up-regulate lipocalin-2 (Lcn2) and investigated whether the engineered MSCs (MSC-Lcn2) could improve cisplatin-induced AKI in a rat model...
June 2, 2017: Cytotechnology
https://www.readbyqxmd.com/read/28551826/development-of-an-8-color-antibody-panel-for-functional-phenotyping-of-human-cd8-cytotoxic-t-cells-from-peripheral-blood-mononuclear-cells
#17
Tara Patel, Amy Cunningham, Martha Holland, John Daley, Suzan Lazo, F Stephen Hodi, Mariano Severgnini
The study of CD8 positive cells in peripheral blood has become an essential part of research in the field of cancer immunotherapies, vaccine development, inflammation, autoimmune disease, etc. In this study, an 8-color flow cytometry panel, containing lineage and functional markers, was developed for the identification of CD8+ cytotoxic T cells in previously cryopreserved peripheral blood mononuclear cells from healthy human donors. By studying functional markers in naïve and CD3/CD28 activated T cells we demonstrate that the panel is capable of detecting protein markers corresponding to different T cell activation statuses...
May 27, 2017: Cytotechnology
https://www.readbyqxmd.com/read/28550492/microrna-889-is-downregulated-by-histone-deacetylase-inhibitors-and-confers-resistance-to-natural-killer-cytotoxicity-in-hepatocellular-carcinoma-cells
#18
Haitao Xie, Qiugui Zhang, Hui Zhou, Jun Zhou, Ji Zhang, Yan Jiang, Jinghong Wang, Xianglin Meng, Leping Zeng, Xiaoxin Jiang
Major histocompatibility complex class I chain-related gene B (MICB) is expressed on tumor cells and participates in natural killer (NK) cell-mediated antitumor immune response through engagement with the NKG2D receptor. This study was undertaken to identify novel microRNA (miRNA) regulators of MICB and clarify their functions in NK cell-mediated cytotoxicity to hepatocellular carcinoma (HCC) cells. Bioinformatic analysis and luciferase reporter assay were conducted to search for MICB-targeting miRNAs. Overexpression and knockdown experiments were performed to determine the roles of candidate miRNAs in the susceptibility of HCC cells to NK lysis...
May 26, 2017: Cytotechnology
https://www.readbyqxmd.com/read/28540540/molecular-mechanisms-of-bombyx-batryticatus-ethanol-extract-inducing-gastric-cancer-sgc-7901-cells-apoptosis
#19
Jin-Yi Wu, Almutamad Sheikho, He Ma, Tian-Ci Li, Ya-Qi Zhao, Ya-Lin Zhang, Dun Wang
Bombyx batryticatus is a traditional Chinese medicine. To understand apoptotic effect of B. batryticatus ethanol extract (BBE), we investigated the role of BBE in inducing apoptosis of human gastric cancer cells SGC-7901. Cells treated with BBE and apoptosis was assessed by methyl thiazolyl tetrazolium (MTT) assay, morphological changes, DNA fragmentation and flow cytometry assays. The expression of Bcl-2, Bax and P21 were evaluated by western blot analysis and real time polymerase chain reaction. MTT assay showed that the cytotoxicity of BBE extract on SGC-7901 cells was correlated with treatment time and concentration...
May 24, 2017: Cytotechnology
https://www.readbyqxmd.com/read/28536871/impact-of-early-subcultures-on-stemness-migration-and-angiogenic-potential-of-adipose-tissue-derived-stem-cells-and-their-resistance-to-in-vitro-ischemic-condition
#20
Hossein Faghih, Arash Javeri, Masoumeh Fakhr Taha
Adipose tissue-derived stem cells (ADSCs) are capable of multipotential differentiation and express several angiogenic, anti-apoptotic and immunomodulatory markers. These features make adipose tissue as a promising source of stem cells for regenerative medicine. However, for efficient translational use, culture-induced changes in the gene expression profile and resistance of the ADSCs to ischemic environment should be taken into consideration. We compared the expression of some clinically important markers between the unpassaged and third-passaged ADSCs by RT-PCR, qPCR and flow cytometry...
May 23, 2017: Cytotechnology
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