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Zhuqiu Xu, Lu Yu, Haibin Lu, Weifeng Feng, Lulu Chen, Jing Zhou, Xiaonan Yang, Zuoliang Qi
We modified an existing protocol to develop a more efficient method to acquire and culture muscle-derived stem cells (MDSCs) and compared the characteristics of cells obtained from the two methods. This method is based on currently used multistep enzymatic digestion and preplate technique. During the replating process, we replaced the traditional medium with isolation medium to promote fibroblast-like cell adherence at initial replating step, which shortened the purifying duration by up to 4 days. Moreover, we modified the culture container to provide a stable microenvironment that promotes MDSC adherence...
November 11, 2018: Cytotechnology
Letícia E Charelli, Nathalia Müller, Karina R Silva, Luís Maurício T R Lima, Celso Sant'Anna, Leandra S Baptista
Stem cell tissue constructs are likely to come into contact with silver-based nanoparticles-such as silver chloride nanoparticles (AgCl-NPs)-used as microbicidals at the implant site or in cosmetics. However, the effect of silver-based nanoparticles on 3D cell cultures with potential for tissue engineering has received little attention. Here, we examined the effect of sub-lethal doses (5, 10 and 25 µg/mL, for 1, 7 and 21 days) of AgCl-NPs produced by 'green' bacterial-based synthesis on spheroid 3D cultures of human adipose tissue stem cells (ASCs)...
November 1, 2018: Cytotechnology
Lenisa Geron, Karina Bezerra Salomão, Kleiton Silva Borges, Augusto Faria Andrade, Carolina Alves Pereira Corrêa, Carlos Alberto Scrideli, Luiz Gonzaga Tone
Medulloblastoma (MB) is the most common malignant childhood brain tumor. MB is currently classified into four molecular subgroups (Wnt, Shh, Group 3, and Group 4). The wingless (Wnt) pathway is responsible for embryonic development and is deregulated in MB. We analyzed the activation of the Wnt pathway in MB cell lines and its correlation with the Shh pathway, with emphasis on the importance of cellular characterization. Transient β-catenin transfection led to an increase in the β-catenin gene and protein expression in MB cell lines...
October 29, 2018: Cytotechnology
Sara Rastgar, Abdolali Movahedinia, Negin Salamat, Amirparviz Salati, Ebrahim Zabihi
The aim of this research was a comparative study on the isolation and culture of head kidney macrophages derived from Acipenser persicous and Rutilus frisii kutum as teleost and chondrostei species of fish. The macrophages were isolated by density gradient sedimentation, followed by adherence to a plastic surface. They exhibited strong phagocytic activity against bacteria. The effect of cell density, incubation time, FBS percentage, pH and temperatures on the cell number and viability were determined and compared...
October 10, 2018: Cytotechnology
Chi-Hsien Liu, Yi-Xin Liu, Wei-Chi Wu
Most bio-industrial mammalian cells are cultured in serum-free media to achieve advantages, such as batch consistency, suspended growth, and simplified purification. The successful development of a serum-free medium could contribute to a reduction in the experimental variation, enhance cell productivity, and facilitate biopharmaceuticals production using the cell culture process. Commercial serum-free media are also becoming more and more popular. However, the cell line secrets its own recombinant product and has special nutritional requirements...
October 3, 2018: Cytotechnology
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
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...
October 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...
October 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...
October 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...
October 2018: Cytotechnology
Kun Liu, Wei Zhang, Yunlong Li, Qingfeng Ding, Yunan Bai, Fachen Wang, Guangming Xu
The potency of mesenchymal stem cells (MSCs) for tissue repair and regeneration is mainly based on their ability to secret beneficial molecules. Administration of MSCs has been proposed as an innovative approach and is proved by a number of clinical trials to a certain degree for the therapy of many diseases including Parkinson's disease (PD). However, the efficacy of MSCs alone is not significant. We investigated the effect of neurotrophic tyrosine receptor kinase 1 (NTRK1) overexpressed peripheral blood MSCs (PB-MSCs) on PD rat model...
October 2018: Cytotechnology
Shiva Mohamadi Moghanjoghi, Meysam Ganjibakhsh, Neda Sadat Gohari, Mehrnaz Izadpanah, Hedieh Rahmati, Zahra Elyasi Gorji, Nazanin Mohebali, Faezeh Vakhshiteh, Parvaneh Farzaneh
Some of lizard species have the ability to lose their tail in order to defend against predators and regenerate the new tail. Lizard's regenerated tail has attracted scientists' attention for unraveling the regeneration process, but less information is known about the cellular characterization and cell growth properties of original tail. This research aimed to report cell culture and banking process of rough-tailed gecko or Cyrtopodion scabrum's original tail cell sample from inner tissue without skin using tissue explant technique...
October 2018: Cytotechnology
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