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Shuichi Kimura, Takeshi Omasa
Chinese hamster ovary (CHO) cells have frequently been used in biotechnology as a mammalian host cell platform for expressing genes of interest. Previously, we constructed a detailed physical chromosomal map of the CHO DG44 cell line by fluorescence in situ hybridization (FISH) imaging using 303 bacterial artificial chromosome (BAC) clones as hybridization probes (BAC-FISH). BAC-FISH results revealed that the two longest chromosomes were completely paired. However, other chromosomes featured partial deletions or rearrangements...
July 9, 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...
July 5, 2018: Cytotechnology
Thouria Bourhim, Myra O Villareal, Chemseddoha Gadhi, Abdellatif Hafidi, Hiroko Isoda
Oil extraction from the kernels of Argania spinosa (L.) Skeels (Sapotaceae), an endemic tree of Morocco, produces argan press-cake (APC) used as a shampoo and to treat sprains, scabies, and for healing wounds. We have previously reported that argan oil has antimelanogenesis effect. Here, we determined if the by-product, APC, has melanogenesis regulatory effect using B16 murine melanoma cells. The effect of APC ethanol extract on cell proliferation and melanin content of B16 cells were measured, and to elucidate the mechanism involved, the expression level of melanogenic enzymes tyrosinase (TYR), dopachrome tautomerase (DCT), and tyrosinase-related protein 1 (TRP1) were determined and mRNA expression level of microphthalmia- associated transcription factor (Mitf) and Tyr genes were quantified...
June 26, 2018: Cytotechnology
Aysel Koç Demir, Ayşe Eser Elçin, Yaşar Murat Elçin
The objective of this study is to evaluate the in vitro and in vivo osteogenic potential of rat bone marrow mesenchymal stem cells (BM-MSCs) using chitosan/hydroxyapatite (C/HAp) microbeads as encapsulation matrix under osteoinductive medium and dynamic culture conditions. The degradation characteristics of C/HAp microbeads were evaluated under in vitro and in vivo conditions for 180 days. BM-MSCs were encapsulated in C/HAp microbeads with > 85% viability, and were cultured in a slow turning lateral vessel-type rotating bioreactor simulating microgravity conditions for 28 days, under the effect of osteogenic inducers...
June 25, 2018: Cytotechnology
Ibrahim Mortada, Rola Mortada
Dental pulp stem cells constitute an attractive source of multipotent mesenchymal stem cells owing to their high proliferation rate and multilineage differentiation potential. Osteogenesis is initiated by osteoblasts, which originate from mesenchymal stem cells. These cells express specific surface antigens that disappear gradually during osteodifferentiation. In parallel, the appearance of characteristic markers, including alkaline phosphatase, collagen type I, osteocalcin and osteopontin characterize the osteoblastic phenotype of dental pulp stem cells...
June 25, 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...
June 8, 2018: Cytotechnology
Renata Mikstacka, Małgorzata Zielińska-Przyjemska, Zbigniew Dutkiewicz, Michał Cichocki, Tomasz Stefański, Mariusz Kaczmarek, Wanda Baer-Dubowska
The aim of this study was to evaluate the cytotoxicity of a series of seven 4'-methylthio-trans-stilbene derivatives against cancer cells: MCF7 and A431 in comparison with non-tumorigenic MCF12A and HaCaT cells. The mechanism of anti-proliferative activity of the most cytotoxic trans-resveratrol analogs: 3,4,5-trimethoxy-4'-methylthio-trans-stilbene (3,4,5-MTS) and 2,4,5-trimethoxy-4'-methylthio-trans-stilbene (2,4,5-MTS) was analyzed and compared with the effect of trans-resveratrol. All the compounds that were studied exerted a stronger cytotoxic effect than trans-resveratrol did...
May 28, 2018: Cytotechnology
Weiliang Feng, Chenlu Liang, Chen Wang, Xingfei Yu, Qinglin Li, Hongjian Yang
To explore the role of ribosomal protein S15A (RPS15A) in breast cancer. The Oncomine database was used to compare the expression of RPS15A in human breast cancer tissues and normal tissues. RPS15A in breast cancer cell line ZR-75-30 and BT474 was specifically knocked down using lentivirus-mediated short hairpin RNAs (shRNAs). RPS15A knockdown efficiency was validated by quantitative polymerase chain reaction and western blot analysis. Subsequently, the functional effects of RPS15A on proliferation of breast cancer cells were investigated by MTT, colony formation and flow cytometry assays...
May 25, 2018: Cytotechnology
Eshtiyag Abdalla Abdalkareem, Ching Yi Ong, Boon Huat Lim, Boon Yin Khoo
The interleukin-21 (IL-21) protein was found to be expressed at an elevated level in clinical samples of colorectal cancer patients without or with a parasitic infection that were collected from Sudan in our previous study. The IL-21 gene in HT29 and HCT116 cells was then correlated to cell proliferation and cell migration, as well as the cellular mechanisms associated with gene expressions in our present study. Our results demonstrated that silencing the IL-21 gene in HCT116 cells increased the cytotoxic level and fibroblast growth factor-4 (FGF4) mRNA expression in the cancer cells...
May 25, 2018: Cytotechnology
Mengkai Yang, Songsong Teng, Chunhui Ma, Yinxian Yu, Peilin Wang, Chengqing Yi
Mesenchymal stem cell (MSC) aging seriously affects its function in stem cell transplantation for treatment. Extensive studies have focused on how to inhibit senescence in MSCs. However, the mechanism of senescence in MSC was not clear. In this study, we used D-galactose to induce MSC aging. Then we found that the number of aging cells was increased compared with untreated MSCs. We discovered that ascorbic acid could inhibit the production of reactive oxygen species (ROS) and activation of AKT/mTOR signaling in MSCs caused by D-galactose...
May 18, 2018: Cytotechnology
Yumei Zhang, Qianqian Wang, Kailing Liu, Enen Gao, Hong Guan, Jian Hou
Incomplete epigenetic reprogramming is one of the major factors affecting the development of embryos cloned by somatic cell nuclear transfer (SCNT). Histone 3 lysine 9 (H3K9) trimethylation has been identified as a key barrier to efficient reprogramming by SCNT. The aim of this study was to explore a method of downregulating H3K9me3 levels in donor cells by using histone lysine demethylase (KDM) protein. When sheep fetal fibroblast cells were treated with recombinant human KDM4D protein (rhKDM4D), the levels of H3K9 trimethylation and dimethylation were both significantly decreased...
May 15, 2018: Cytotechnology
Roberto Carrio, Ge Zhang, Donald R Drake, Brian C Schanen
Although a variety of assays have been used to examine T cell responses in vitro, standardized ex vivo detection of antigen-specific CD4+ T cells from human circulatory PBMCs remains constrained by low-dimensional characterization outputs and the need for polyclonal, mitogen-induced expansion methods to generate detectable response signals. To overcome these limitations, we developed a novel methodology utilizing antigen-pulsed autologous human dendritic target cells in a rapid and sensitive assay to accurately enumerate antigen-specific CD4+ T cell precursor frequency by multiparametric flow cytometry...
May 7, 2018: Cytotechnology
K Jurica, I Brčić Karačonji, A Mikolić, D Milojković-Opsenica, V Benković, N Kopjar
Strawberry tree (Arbutus unedo L.) leaves have long been used in the traditional medicine of the Mediterranean region. One of their most bioactive constituents is the glycoside arbutin, whose presence makes A. unedo suitable as a potential substitute for bearberry [Arctostaphylos uva ursi (L.) Spreng] leaves, an herbal preparation widely used for treating urinary tract infections. The safety and biocompatibility of strawberry tree water leaf extract have not yet been documented well. This study estimated arbutin content in strawberry tree water leaf extract (STE) using high performance liquid chromatography...
April 25, 2018: Cytotechnology
Zhao Chong, Haruka Matsuo, Mai Kuroda, Shuntaro Yamashita, Gopal Prasad Parajuli, Hira Kaji Manandhar, Kuniyoshi Shimizu, Yoshinori Katakura
Mushrooms possess various bioactivities and are used as nutritional supplements and medicinal products. Twenty-nine bioactive components have been extracted recently from mushrooms grown in Nepal. In this study, we evaluated the ability of these mushroom extracts to augment SIRT1, a mammalian SIR2 homologue localized in cytosol and nuclei. We established a system for screening food ingredients that augment the SIRT1 promoter in HaCaT cells, and identified a SIRT1-augmenting mushroom extract (number 28, Trametes versicolor)...
June 2018: Cytotechnology
Ayane Hori, Takashi Fujimura, Seiji Kawamoto
An intravenous injection of plasma-derived immunoglobulins is used for the treatment of severe infectious and autoimmune disorders. Despite of its clinical efficacy, precise mechanisms by which intravenous immunoglobulin (IVIg) suppresses proinflammatory immune response are still enigmatic. Here, we provide in vitro evidence that IVIg inhibits homeostatic proliferation of B cells accompanied by induction of their cell aggregation. The IVIg-driven suppression of B cell proliferation and induction of cell aggregation are both unaffected by treatment with a neutralizing antibody against low-affinity Fc receptors for IgG (CD16/FcγRIII and CD32/FcγRII), known cell surface ligands for IVIg...
June 2018: Cytotechnology
Gaoxiong Wang, Youshi Zheng, Yingchao Wang, Zhixiong Cai, Naishun Liao, Jingfeng Liu, Wenmin Zhang
Although hepatocyte transplantation and bioartificial liver support system provide new promising opportunities for those patients waiting for liver transplantation, hepatocytes are easily losing liver-specific functions by using the common in vitro cultured methods. The co-culture strategies with mimicking the in vivo microenvironment would facilitate the maintenance of liver-specific functions of hepatocytes. Considering that hepatocytes and endothelial cells (ECs) account for 80-90% of total cell populations in the liver, hepatocytes and ECs were directly co-cultured with hepatic stellate cells (HSCs) or adipose tissue-derived stem cells (ADSCs) at a ratio of 700:150:3 or 14:3:3 in the present study, and the liver-specific functions were carefully analyzed...
April 19, 2018: Cytotechnology
Zhong-Bao Ruan, Ge-Cai Chen, Yin Ren, Li Zhu
We aimed to investigate the differentially expressed long non-coding RNAs (lncRNAs) during the differentiation of human umbilical cord derived mesenchymal stem cells (hUCMSCs) into cardiomyocyte-like cells induced by 5-aza. hUCMSCs were isolated and purified from umbilical cords. After treated with 10 μmol/L 5-Aza for 24 h, hUCMSCs wereas continued to be cultured for 14 days. Comparison of cardiac specific genes and the expression profile of lncRNAs on hUCMSCs between day 14 and day 0 was performed using immunofluorescence staining, immunohistochemistry, Western blot assay, RT-PCR and lncRNA microarray...
April 9, 2018: Cytotechnology
Zhi-Gang Liu, Wei-Feng Ding, Shi-Cong Xie, Na Sun, Xin Zhang, Xian Li, Ying Feng
Cell cloning is of great importance in keeping particular properties of cultured cells, and interesting cells can be selected by cloning from heterogeneous cell populations. In addition, continuous cell lines usually from primary culture are prone to heterologous constitution and genetic instability, so that supplementary cloning steps are necessary for achieving a homogenous cell population. In this study, limiting dilution culture and feeder layer culture were originally used for cloning RIRI-PaDe-3 cell line, but both failed...
April 9, 2018: Cytotechnology
Li Jing, Wang Haitao, Wang Qiong, Zhu Fu, Zhang Nan, Zhang Xuezheng
Early diagnosis and changes associated with atherosclerosis are crucial in clinical medicine. However, atherosclerosis is a multifactorial disease. Asiaticoside (AA), a triterpenoid derived from Centella asiatica, has anti-inflammatory activity. Endothelium-derived nitric oxide is important in modulating vascular tone in a distinct vessel size-dependent manner; it plays a dominant role in conduit arteries and endothelium-dependent hyperpolarisation in resistance vessels. This study evaluated the effects of AA administration on human umbilical endothelial cells with oxidised low-density lipoprotein-induced inflammation...
April 2018: Cytotechnology
Somaieh Zafari, Mohsen Sharifi, Najmeh Ahmadian Chashmi
The callus and hairy root cultures of Prosopis farcta were established to develop effective strategies to enhance its valuable and medicinally important flavonoid compounds. For callus induction, the hypocotyl, cotyledon and shoot explants were subjected to different plant hormones, naphthalene acetic acid (NAA), benzylaminopurine (BAP), kinetin and dichlorophenoxyacetic acid (2,4-D). Greater callus induction was obtained from hypocotyl explants on MS medium containing 3.0 mg L-1 NAA + 2.0 mg L-1 BAP...
April 2018: Cytotechnology
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