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Katja Rebolj, Matija Veber, Matej Drobnič, Elvira Maličev
Mesenchymal stem cells (MSCs) are heterogeneous population of cells with great potential for regenerative medicine. MSCs are relatively easy to expand in a cell culture, however determination of their concentration in harvested tissue is more complex and is not implemented as routine procedure. To identify MSCs collected from bone marrow we have used two combinations of cell markers (CD45- /CD73+ /CD90+ /CD105+ and CD45- /CD271+ ) and fibroblast colony-forming unit (CFU-F) assay. Further, in donors of various ages, mesenchymal stem cell concentration was compared with the result of CFU-F assay and with hematopoietic stem cell concentration, determined by a standardized flow cytometric assay...
September 18, 2018: Cytotechnology
Ryo Gouko, Manabu Onuma, Takahiro Eitsuka, Masafumi Katayama, Kouhei Takahashi, Kiyotaka Nakagawa, Miho Inoue-Murayama, Tohru Kiyono, Tomokazu Fukuda
Tsushima leopard cat is the subspecies of Amur cats, and it is classified as the most highest class of critically endangered animals. Although the protection activity is highly recognized, the number of animals is decreasing due to the human activity and invasion of domestic cats and infectious disease. In this study, we succeeded primary culture of normal fibroblasts derived from the Tsushima leopard cat (Prionailurus bengalensis euptilurus). Furthermore, we introduced the human derived mutant Cyclin Dependent Kinase 4, Cyclin D1, and telomere reverse transcriptase...
September 17, 2018: Cytotechnology
Mateus Eugenio, Loraine Campanati, Nathalia Müller, Luciana F Romão, Jorge de Souza, Soniza Alves-Leon, Wanderley de Souza, Celso Sant'Anna
Glioblastomas (GBM) are aggressive brain tumors with very poor prognosis. While silver nanoparticles represent a potential new strategy for anticancer therapy, the silver/silver chloride nanoparticles (Ag/AgCl-NPs) have microbicidal activity, but had not been tested against tumor cells. Here, we analyzed the effect of biogenically produced Ag/AgCl-NPs (from yeast cultures) on the proliferation of GBM02 glioblastoma cells (and of human astrocytes) by automated, image-based high-content analysis (HCA). We compared the effect of 0...
September 10, 2018: Cytotechnology
Fieke W Hoff, Chenyue W Hu, Amina A Qutub, Yihua Qiu, Elizabeth Graver, Giang Hoang, Manasi Chauhan, Eveline S J M de Bont, Steven M Kornblau
Mycoplasma contamination is a major problem in cell culturing, potentially altering the results of cell line-based experiments in largely uncharacterized ways. To define the consequences of mycoplasma infection at the level of protein expression we utilized the reverse phase protein array technology to analyze the expression of 235 proteins in mycoplasma infected, uninfected post treatment, and never-infected leukemic cell lines. Overall, protein profiles of cultured cells remained relatively stable after mycoplasma infection...
September 6, 2018: Cytotechnology
Ya Li, Weifeng Zhang, Junli Zhao, Sai Li, Linlin Shan, Jiuling Zhu, Yan Li, He Zhu, Qinwen Mao, Haibin Xia
The clustered regulatory interspersed short palindromic repeat (CRISPR)/CRISPR-associated protein (Cas) system has been widely used for gene knock-out. Lentiviral vectors have been commonly used as a delivery method for this system, however, prolonged Cas9/sgRNA expression due to lentiviral integration can lead to accumulating off-target mutations. To solve this issue in engineering a gene knock-out cell line, this study established a novel system, which was composed of two lentiviral vectors. One lentiviral vector carried simultaneously sgRNAs and CRISPR/Cas9 expression cassettes targeting single or multiple gene(s); the other lentiviral vector carried Cre that could remove excess sgRNAs and Cas9 expression cassettes in the genome after gene targeting was achieved...
September 1, 2018: Cytotechnology
Ewa Skała, Monika Toma, Tomasz Kowalczyk, Tomasz Śliwiński, Przemysław Sitarek
Rhaponticum carthamoides transformed root extract induces double strand DNA damage by increasing the number of phosphorylated H2A.X- and cleaved PARP1-positive U87MG cells and patient-derived IV grade glioma cells. Furthermore, treatment of these cells with root extract causes down-regulation of UHRF1 and DNMT1. Transformed root extract is rich in caffeoylquinic acid derivatives, especially tricaffeoylquinic acid derivatives. Our findings demonstrate that the R. carthamoides transformed root extract may trigger apoptosis in glioma cells by induction of DNA damage, PARP cleavage and epigenetic modification...
August 31, 2018: Cytotechnology
Rashad Mogalli, Toshiya Matsukawa, Osamu Shimomura, Hiroko Isoda, Nobuhiro Ohkohchi
Mitochondrial dysfunction has been identified as one of the primary factors contributing to liver diseases. Pathways that control mitochondrial biogenesis are potential therapeutic targets for the amelioration of hepatocyte dysfunction and liver disease. Research on natural pharmacological agents that ameliorate liver diseases has intensified over the last two decades. Cyanidin-3-glucoside (Cy3g), a dietary flavonoid compound extracted from a wide variety of fruits and vegetables, reportedly has several beneficial health effects...
August 28, 2018: Cytotechnology
Gakuro Harada, Shiori Onoue, Chisato Inoue, Sakae Hanada, Yoshinori Katakura
Delphinidin is an anthocyanidin commonly found in various fruits and vegetables. Delphinidin has been known to possess many functions, such as an antioxidant, and anti-inflammatory, anti-cancer and anti-muscular atrophy agent. In this study, we attempted to evaluate the effects of delphinidin on lipid accumulation in hepatocytes. The results showed that palmitic acid (PA)-induced cellular senescence in HepG2 cells and reduced the expression of SMARCD1, which is known to regulate senescence-associated lipid accumulation in hepatocytes...
August 28, 2018: Cytotechnology
Mehmet Varol
Natural products have been used for centuries as the most potent remedies to cure many diseases including cancer diseases. Angiogenesis is defined as the formation of new capillaries from existing vessels and plays a key role in the tumorigenesis process. Barbatolic acid is a little known lichen-derived small-molecule. In the present study, barbatolic acid was isolated from the acetone extract of Bryoria capillaris, and its anti-breast cancer and anti-angiogenic potential was investigated using human umbilical vein endothelial cells (HUVECs), human breast ductal carcinoma (T-47D) and cisplatin-resistant BRCA2-mutated human breast TNM stage IV adenocarcinoma (HCC1428) cells...
August 28, 2018: Cytotechnology
Gönül Ulus, A Tansu Koparal, Kemal Baysal, Günay Yetik Anacak, N Ülkü Karabay Yavaşoğlu
Cotton, a staple fiber that grows around the seeds of the cotton plants (Gossypium), is produced throughout the world, and its by products, such as cotton fibers, cotton-seed oil, and cottonseed proteins, have a variety of applications. Cotton-seed contains gossypol, a natural phenol compound. (±)-Gossypol is a yellowish polyphenol that is derived from different parts of the cotton plant and contains potent anticancer properties. Tumor growth and metastasis are mainly related to angiogenesis; therefore, anti-angiogenic therapy targets the new blood vessels that provide oxygen and nutrients to actively proliferating tumor cells...
August 19, 2018: Cytotechnology
Guo-Qiang Zheng, Hai-Yu Ji, Shao-Jing Zhang, Juan Yu, An-Jun Liu
In this study, the cytotoxic activity of selenious-β-lactoglobulin (Se-β-Lg) and the anticancer mechanism were investigated in human lung cancer A549 cells in vitro. MTT assay showed that Se-β-Lg at 200 μg/mL exhibited a significant suppression effect on A549 cells and the maximum inhibition rate reached 90% after 72 h treatment. Flow cytometry analysis revealed that 200 μg/mL of Se-β-Lg induced cell cycle arrest at G0 /G1 phase. Cell apoptosis was induced via the generation of reactive oxygen species (ROS) and the decrease of mitochondrial membrane potential (ΔΨm) in a time-dependent manner...
August 10, 2018: Cytotechnology
Mandana Kazem-Arki, Mahboubeh Kabiri, Iman Rad, Nasim Hayati Roodbari, Hoorieh Hosseinpoor, Samaneh Mirzaei, Kazem Parivar, Hana Hanaee-Ahvaz
Recent developments in bone tissue engineering have paved the way for more efficient and cost-effective strategies. Additionally, utilization of autologous sources has been considered very desirable and is increasingly growing. Recently, activated platelet rich plasma (PRP) has been widely used in the field of bone tissue engineering, since it harbours a huge number of growth factors that can enhance osteogenesis and bone regeneration. In the present study, the osteogenic effects of PRP coated nanofibrous PES/PVA scaffolds on adipose-derived mesenchymal stem cells have been investigated...
August 6, 2018: Cytotechnology
Deniz Mortazavi, Mohammadreza Sharifi
New achievements in the field of cancer treatment are results of recent advances in molecular medicine and gene therapy. Usage of microRNAs (miRNAs) which are small noncoding RNAs is one of the molecular research lines for the diagnosis and treatment of cancers. miRNAs have an important role in post-transcriptional regulation of the gene expression and are involved in cellular activities such as growth, differentiation, cell death and cancer development. One of the miRNAs that showed downregulation in human acute erythroleukemia is hsa-let-7c-5p...
August 2, 2018: Cytotechnology
Noriko Yamano, Takeshi Omasa
Chinese hamster ovary cell lines are good manufacturing practice-certified host cells and are widely used in the field of biotechnology to produce therapeutic antibodies. Recombinant protein productivity in cells is strongly associated with cell growth. To control cell proliferation, many approaches have previously been tested including: genetic engineering, chemical additives such as cell cycle inhibitors, and temperature shift of the culture. To be widely adopted in the biopharmaceutical industry, the culture methods should be simple, uniform and safe...
August 1, 2018: Cytotechnology
Swapnil Ganesh Sanmukh, Sérgio Luis Felisbino
This work presents a pipette tip gap closure migration assay prototype tool (semi-adherent relative upsurge-s-ARU-method) to study cell migration or wound healing in semi-adherent cell lines, such as lymph node carcinoma of the prostate (LNCaP). Basically, it consists of a 6-well cover plate modification, where pipette tips with the filter are shortened and fixed vertically to the inner surface of the cover plate, with their heights adjusted to touch the bottom of the well center. This provides a barrier for the inoculated cells to grow on, creating a cell-free gap...
August 1, 2018: Cytotechnology
Narumi Uno, Tomohito Fujimoto, Shinya Komoto, Gene Kurosawa, Masaaki Sawa, Teruhiko Suzuki, Yasuhiro Kazuki, Mitsuo Oshimura
G protein-coupled receptors (GPCRs) are seven-transmembrane domain receptors that interact with the β-arrestin family, particularly β-arrestin 1 (ARRB1). GPCRs interact with 33% of small molecule drugs. Ligand screening is promising for drug discovery concerning GPCR-related diseases. Luciferase complementation assay (LCA) enables detection of protein-protein complementation via bioluminescence following complementation of N- and C-terminal luciferase fragments (NEluc and CEluc) fused to target proteins, but it is necessary to co-express the two genes...
July 31, 2018: Cytotechnology
De-Bin Guo, Xiang-Qing Zhu, Qing-Qing Li, Gao-Mi-Yang Liu, Guang-Ping Ruan, Rong-Qing Pang, Yu-Hao Chen, Qiang Wang, Jin-Xiang Wang, Ju-Fen Liu, Qiang Chen, Xing-Hua Pan
Umbilical cord mesenchymal stem cells (UC-MSCs) exert strong immunomodulatory effects and can repair organs. However, their roles in radiation injury remain unclear. We show that in tree shrews with acute radiation injury, injected UC-MSCs significantly improved survival rates, reduced lung inflammation and apoptosis, prevented pulmonary fibrotic processes, recovered hematopoiesis, and increased blood counts. A protein microarray analysis showed that serum levels of the anti-inflammatory cytokines IL-10 and IL-13 and the growth factors BMP-5, BMP-7, HGF, insulin, NT-4, VEGFR3, and SCF were significantly higher, while those of the inflammatory cytokines IL-2, TIMP-2, TNF-α, IFN-γ, IL-1ra, and IL-8 and the fibrosis-related factors PDGF-BB, PDGF-AA, TGF-β1, IGFBP-2, and IGFBP-4 were significantly lower in UC-MSC-injected animals...
July 31, 2018: Cytotechnology
Kyan Thelen, Stephanie W Watts, G Andres Contreras
Perivascular adipose tissue (PVAT) has the capacity to secrete vasoactive mediators with the potential to regulate vascular function. Given its location adjacent to the vasculature, PVAT dysfunction may be part of the pathophysiology of cardiovascular diseases. To study the mechanisms of PVAT dysfunction, several adipogenic models have been proposed. However, these approaches do not adequately reflect PVAT adipocyte phenotypes variability that depends on their anatomical location. Despite PVAT importance in modulating vascular function, to date, there is not a depot-specific adipogenic model for PVAT adipocytes...
July 27, 2018: Cytotechnology
Yongfeng Zhang, Li Li, Qin Wang, Siyuan Zhan, Linjie Wang, Tao Zhong, Jiazhong Guo, Hongping Zhang
Fibroblast growth factor 21 (FGF21) potentially regulates glucose and lipid metabolism in energy homeostasis. We investigated dynamic changes in goat adipocytes treated with 75 nM FGF21 for 24, 36 and 48 h. Compared to controls, FGF21-treated adipocytes displayed smaller lipid droplets and altered levels of the mRNA transcripts encoding several lipolysis genes. The genes with elevated mRNA levels included: ATGL, HSL, CPT-1, and UCP1, and this was observed mainly at 24 and 36 h (P < 0.05). Some gene expression was attenuated including lipogenesis genes, such as SREBP1, PPARγ, C/EBPα, and ACC...
July 26, 2018: Cytotechnology
Yanxia Luo, Ali Mohsin, Chenze Xu, Qizheng Wang, Haifeng Hang, Yingping Zhuang, Ju Chu, Meijin Guo
The proliferation and migration of mesenchymal stem cells (MSCs) are the efficiency determinants in MSCs transplant therapy. Sertoli cells considered as "nurse cell" possesses the ability to enhance the proliferation and migration of umbilical cord mesenchymal stem cells (UCMSCs). However, no reports about TM4 cells' effect on the proliferation and migration of adipose tissue-derived mesenchymal stem cells (ADSCs) have been found until at present research work. Therefore, this study investigates the effect of TM4 cells on the proliferation and migration of ADSCs...
July 21, 2018: Cytotechnology
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