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https://www.readbyqxmd.com/read/29150045/spectrum-of-benzo-a-pyrene-induced-mutations-in-the-pig-a-gene-of-l5178ytk-cells-identified-with-next-generation-sequencing
#1
Javier Revollo, Yiying Wang, Page McKinzie, Azra Dad, Mason Pearce, Robert H Heflich, Vasily N Dobrovolsky
We used Sanger sequencing and next generation sequencing (NGS) for analysis of mutations in the endogenous X-linked Pig-a gene of clonally expanded L5178YTk(+/-) cells. The clones developed from single cells that were sorted on a flow cytometer based upon the expression pattern of the GPI-anchored marker, CD90, on their surface. CD90-deficient and CD90-proficient cells were sorted from untreated cultures and CD90-deficient cells were sorted from cultures treated with benzo[a]pyrene (B[a]P). Pig-a mutations were identified in all clones developed from CD90-deficient cells; no Pig-a mutations were found in clones of CD90-proficient cells...
December 2017: Mutation Research
https://www.readbyqxmd.com/read/29142448/expression-of-the-bone-morphogenetic-protein-2-bmp2-in-the-human-cumulus-cells-as-a-biomarker-of-oocytes-and-embryo-quality
#2
Sirin B Demiray, Ozlem Yilmaz, Ege N T Goker, Erol Tavmergen, Nilufer Calimlioglu, Ugur Sezerman, Huseyin O Soykam, Gulperi Oktem
Background: The members of the transforming growth factor-B superfamily, as the bone morphogenetic proteins (BMPs) subfamily and anti-Müllerian hormone (AMH), play a role during follicular development, and the bone morphogenetic protein-2 (BMP2), AMH, and THY1 are expressed in ovaries. Aim: This study was designed to define whether or not the expressions of these proteins in human cumulus cells (CCs) can be used as predictors of the oocyte and embryo competence...
July 2017: Journal of Human Reproductive Sciences
https://www.readbyqxmd.com/read/29138638/tissue-source-and-cell-expansion-condition-influence-phenotypic-changes-of-adipose-derived-stem-cells
#3
Lauren H Mangum, Shanmugasundaram Natesan, Randolph Stone, Nicole L Wrice, David A Larson, Kyle F Florell, Barbara A Christy, Maryanne C Herzig, Andrew P Cap, Robert J Christy
Stem cells derived from the subcutaneous adipose tissue of debrided burned skin represent an appealing source of adipose-derived stem cells (ASCs) for regenerative medicine. Traditional tissue culture uses fetal bovine serum (FBS), which complicates utilization of ASCs in human medicine. Human platelet lysate (hPL) is one potential xeno-free, alternative supplement for use in ASC culture. In this study, adipogenic and osteogenic differentiation in media supplemented with 10% FBS or 10% hPL was compared in human ASCs derived from abdominoplasty (HAP) or from adipose associated with debrided burned skin (BH)...
2017: Stem Cells International
https://www.readbyqxmd.com/read/29134714/identification-of-perivascular-and-stromal-mesenchymal-stem-progenitor-cells-in-porcine-endometrium
#4
J Wiater, M Niedziela, A Posmysz, K Wartalski, B Gajda, Z Smorąg, Z Rajfur, J Karasiński
Mammalian uterus contains a population of mesenchymal stem/progenitor cells that likely contribute to endometrial regeneration during each reproductive cycle. In human and mouse, they reside in perivascular, epithelial and stromal compartments of the endometrial functionalis and basalis. Here, we aimed to identify tissue resident cells expressing mesenchymal stem cell markers CD29, CD44, CD90, CD105, CD140b and CD146 in the porcine endometrium. We used single immunofluorescence and Western blotting. Each of these markers was detected in small cells surrounding endometrial blood vessels...
November 14, 2017: Reproduction in Domestic Animals, Zuchthygiene
https://www.readbyqxmd.com/read/29129620/human-adipose-derived-stem-cells-adsc-and-human-periodontal-ligament-stem-cells-pdlsc-as-cellular-substrates-of-a-toxicity-prediction-assay
#5
Natássia Caroline Resende Corrêa, Crisciele Kuligovski, Ariane Caroline Campos Paschoal, Ana Paula Ressetti Abud, Carmen Lucia Kuniyoshi Rebelatto, Lidiane Maria Boldrini Leite, Alexandra Cristina Senegaglia, Bruno Dallagiovanna, Alessandra Melo de Aguiar
With the increasing need to develop in vitro assays to replace animal use, human stem cell-derived methods are emerging and showing outstanding contributions to the toxicological screening of substances. Adult human stem cells such as adipose-derived stem cells (ADSC) and periodontal ligament stem cells (PDLSC) were used as cell substrates for a cytotoxicity assay and toxicity prediction using the neutral red uptake (NRU) assay. First, primary cell cultures from three independent donors, from each tissue source, were characterized as mesenchymal stem cells (MSC) by plastic adherence and appropriate immunophenotype for MSC markers (positive for CD90, CD73, and CD105 and negative for CD11b, CD34, CD45, HLADR, and CD19)...
November 9, 2017: Regulatory Toxicology and Pharmacology: RTP
https://www.readbyqxmd.com/read/29119116/use-of-limiting-dilution-method-for-isolation-of-nucleus-pulposus-mesenchymal-stem-progenitor-cells-and-effects-of-plating-density-on-biological-characteristics-and-plasticity
#6
Linghan Lin, Zhiwei Jia, Yachao Zhao, Yaohong Wu, Xiyan Zhao, Yong Li, Ziming Guo, Jiahai Chen, Shi Cheng, Deli Wang, Dike Ruan
Objectives: To evaluate the effects of the limiting dilution method and plating density in rat nucleus pulposus mesenchymal stem/progenitor cells (NPMSCs). Materials and Methods: Nucleus pulposus tissues were isolated from 12-week-old male Sprague-Dawley rats and NPMSCs were isolated using limiting dilution method. Cells were then classified into 3 groups according to plating density. Cell morphologies were observed, and colony-forming units, migration abilities, proliferative capacities, cell cycle percentages, multilineage differentiation capacities, stem cell biomarker expression levels, and immunophenotyping were also examined in each group...
2017: BioMed Research International
https://www.readbyqxmd.com/read/29118937/isolation-differentiation-and-characterization-of-mesenchymal-stem-cells-from-human-bone-marrow
#7
Kaveh Baghaei, Seyed Mohmoud Hashemi, Samaneh Tokhanbigli, Ali Asadi Rad, Hamid Assadzadeh-Aghdaei, Abdolhamid Sharifian, Mohammad Reza Zali
Aim: We describe the minimum requirements and a simplified method for isolation and characterization of mesenchymal stem cells (MSCs) from human bone marrow. Background: MSCs are well known adult stem cells present in many tissues such as adipocytes, chondrocytes, osteoblasts, and neurons. Many isolations and characterization methods have emerged to apply MSCs in the clinical applications, which many of them are expensive and time-consuming. Methods: MSC isolation was carried out from human bone marrow, and cultured in defined medium...
2017: Gastroenterology and Hepatology From Bed to Bench
https://www.readbyqxmd.com/read/29115020/glycosylphosphatidylinositol-gpi-anchored-protein-deficiency-serves-as-a-reliable-reporter-of-pig-a-gene-mutation-support-from-an-in-vitro-assay-based-on-l5178y-tk-cells-and-the-cd90-2-antigen
#8
Jeffrey C Bemis, Svetlana L Avlasevich, Carson Labash, Page McKinzie, Javier Revollo, Vasily N Dobrovolsky, Stephen D Dertinger
Lack of cell surface glycosylphosphatidylinositol (GPI)-anchored protein(s) has been used as a reporter of Pig-a gene mutation in several model systems. As an extension of this work, our laboratory initiated development of an in vitro mutation assay based on the flow cytometric assessment of CD90.2 expression on the cell surface of the mouse lymphoma cell line L5178Y/Tk(+/-) . Cells were exposed to mutagenic and nonmutagenic compounds for 24 hr followed by washout and incubation for an additional 7 days. Following this mutant manifestation time, cells were labeled with fluorescent antibodies against CD90...
November 8, 2017: Environmental and Molecular Mutagenesis
https://www.readbyqxmd.com/read/29105402/adipose-derived-stem-cells-affect-mir-145-and-p53-expressions-of-co-cultured-hematopoietic-stem-cells
#9
Tahereh Foroutan, Aisan Farhadi, Saeed Abroun, Bahram Mohammad Soltani
OBJECTIVES: Umbilical cord blood is used for transplantation purposes in regenerative medicine of hematological disorders. MicroRNAs are important regulators of gene expression that control both physiological and pathological processes such as cancer development and incidence. There is a new relation between p53 (tumor suppressor gene) and miR-145 (suppressor of cell growth) upregulation. In this study, we have assessed how adipose-derived stem cells (ADSCs) affect the expansion of hematopoietic stem cells (HSCs), as well as miR-145 and p53 expressions...
January 2018: Cell Journal
https://www.readbyqxmd.com/read/29105086/dose-dependent-effect-of-triiodothyronine-on-the-chondrogenic-differentiation-of-mesenchymal-stem-cells-from-the-bone-marrow-of-female-rats
#10
Higor A Assis, Nathalia C Elert, André Luiz B P Azevedo, Iolanda S Braga, Rogéria Serakides, Natália de Melo Ocarino, Alfredo M de Goes, Francisco de Paula Careta, Greiciane G Paneto, Adriana M A da Silva, Jankerle N Boeloni
OBJECTIVES: Verify the in-vitro effect of triiodothyronine (T3) on the chondrogenic differentiation of female rat bone marrow mesenchymal stem cells (BMMSCs) over several time periods and at several doses. METHODS: CD54 + /CD73 + /CD90 +  BMMSCs from Wistar female rats were cultured in chondrogenic medium with or without T3 (0.01; 1; 100; 1000 nm). At seven, 14 and 21 days, the cell morphology, chondrogenic matrix formation and expression of Sox9 and collagen II were evaluated...
November 3, 2017: Journal of Pharmacy and Pharmacology
https://www.readbyqxmd.com/read/29103027/inhibition-of-mir-155-a-therapeutic-target-for-breast-cancer-prevented-in-cancer-stem-cell-formation
#11
Jiangcheng Zuo, Yalan Yu, Man Zhu, Wei Jing, Mingxia Yu, Hongyan Chai, Chunzi Liang, Jiancheng Tu
BACKGROUND: Breast cancer is a common cancer in women of worldwide. Cancer cells with stem-like properties played important roles in breast cancer, such as relapse, metastasis and treatment resistance. Micro-RNA-155 (miR-155) is a well-known oncogenic miRNA overexpressed in many human cancers. METHODS: The expression levels of miR-155 in 38 pairs of cancer tissues and adjacent normal tissues from breast cancer patients were detected using quantitative real-time PCR...
October 27, 2017: Cancer Biomarkers: Section A of Disease Markers
https://www.readbyqxmd.com/read/29098723/establishing-a-novel-pig-a-gene-mutation-assay-in-l5178ytk-mouse-lymphoma-cells
#12
Yiying Wang, Javier Revollo, Page McKinzie, Mason G Pearce, Azra Dad, Berran Yucesoy, Hans Rosenfeldt, Robert H Heflich, Vasily N Dobrovolsky
The X-linked Pig-a gene encodes an enzyme required for the biosynthesis of glycosyl phosphatidylinositol (GPI) anchors. Pig-a mutant cells fail to synthesize GPI and to express GPI-anchored protein markers (e.g., CD90) on their surface. Marker deficiency serves as a phenotypic indicator of Pig-a mutation in various in vivo assays. Here, we describe an in vitro Pig-a mutation assay in L5178YTk(+/-) mouse lymphoma cells, in which mutant-phenotype cells are measured by flow cytometry using a fluorescent anti-CD90 antibody...
November 2, 2017: Environmental and Molecular Mutagenesis
https://www.readbyqxmd.com/read/29097870/adipose-derived-stromal-cells-resemble-bone-marrow-stromal-cells-in-hepatocyte-differentiation-potential-in-vitro-and-in-vivo
#13
Li-Juan Xu, Shu-Fang Wang, De-Qing Wang, Lian-Jun Ma, Zheng Chen, Qian-Qian Chen, Jun Wang, Li Yan
AIM: To investigate whether mesenchymal stem cells (MSCs) from adipose-derived stromal cells (ADSCs) and bone marrow stromal cells (BMSCs) have similar hepatic differentiation potential. METHODS: Mouse ADSCs and BMSCs were isolated and cultured. Their morphological and phenotypic characteristics, as well as their multiple differentiation capacity were compared. A new culture system was established to induce ADSCs and BMSCs into functional hepatocytes. Reverse transcription polymerase chain reaction, Western blot, and immunofluorescence analyses were performed to identify the induced hepatocyte-like cells...
October 14, 2017: World Journal of Gastroenterology: WJG
https://www.readbyqxmd.com/read/29096714/isolation-and-prolonged-expansion-of-oral-mesenchymal-stem-cells-under-clinical-grade-gmp-compliant-conditions-differentially-affects-stemness-properties
#14
Athina Bakopoulou, Danae Apatzidou, Eleni Aggelidou, Evangelia Gousopoulou, Gabriele Leyhausen, Joachim Volk, Aristeidis Kritis, Petros Koidis, Werner Geurtsen
BACKGROUND: Development of clinical-grade cell preparations is central to meeting the regulatory requirements for cellular therapies under good manufacturing practice-compliant (cGMP) conditions. Since addition of animal serum in culture media may compromise safe and efficient expansion of mesenchymal stem cells (MSCs) for clinical use, this study aimed to investigate the potential of two serum/xeno-free, cGMP culture systems to maintain long-term "stemness" of oral MSCs (dental pulp stem cells (DPSCs) and alveolar bone marrow MSCs (aBMMSCs)), compared to conventional serum-based expansion...
November 2, 2017: Stem Cell Research & Therapy
https://www.readbyqxmd.com/read/29094769/differentiation-and-characterization-of-rat-adipose-tissue-mesenchymal-stem-cells-into-endothelial-like-cells
#15
V Cannella, G Piccione, R Altomare, A Marino, P Di Marco, L Russotto, S Di Bella, G Purpari, F Gucciardi, G Cassata, G Damiano, V D Palumbo, A Santoro, C Russo Lacerna, A I Lo Monte, A Guercio
In this study, mesenchymal stem cells were isolated from rat adipose tissue (AD-MSCs) to characterize and differentiate them into endothelial-like cells. AD-MSCs were isolated by mechanical and enzymatic treatments, and their identity was verified by colony-forming units (CFU) test and by differentiation into cells of mesodermal lineages. The endothelial differentiation was induced by plating another aliquot of cells in EGM-2 medium, enriched with specific endothelial growth factors. Five subcultures were performed...
November 2, 2017: Anatomia, Histologia, Embryologia
https://www.readbyqxmd.com/read/29093179/a-distinct-hematopoietic-stem-cell-population-for-rapid-multilineage-engraftment-in-nonhuman-primates
#16
Stefan Radtke, Jennifer E Adair, Morgan A Giese, Yan-Yi Chan, Zachary K Norgaard, Mark Enstrom, Kevin G Haworth, Lauren E Schefter, Hans-Peter Kiem
Hematopoietic reconstitution after bone marrow transplantation is thought to be driven by committed multipotent progenitor cells followed by long-term engrafting hematopoietic stem cells (HSCs). We observed a population of early-engrafting cells displaying HSC-like behavior, which persisted long-term in vivo in an autologous myeloablative transplant model in nonhuman primates. To identify this population, we characterized the phenotype and function of defined nonhuman primate hematopoietic stem and progenitor cell (HSPC) subsets and compared these to human HSPCs...
November 1, 2017: Science Translational Medicine
https://www.readbyqxmd.com/read/29078802/restoring-the-quantity-and-quality-of-elderly-human-mesenchymal-stem-cells-for-autologous-cell-based-therapies
#17
Travis J Block, Milos Marinkovic, Olivia N Tran, Aaron O Gonzalez, Amanda Marshall, David D Dean, Xiao-Dong Chen
BACKGROUND: Degenerative diseases are a major public health concern for the aging population and mesenchymal stem cells (MSCs) have great potential for treating many of these diseases. However, the quantity and quality of MSCs declines with aging, limiting the potential efficacy of autologous MSCs for treating the elderly population. METHODS: Human bone marrow (BM)-derived MSCs from young and elderly donors were obtained and characterized using standard cell surface marker criteria (CD73, CD90, CD105) as recommended by the International Society for Cellular Therapy (ISCT)...
October 27, 2017: Stem Cell Research & Therapy
https://www.readbyqxmd.com/read/29073604/cardiac-mesenchymal-stem-cells-proliferate-early-in-the-ischemic-heart
#18
Christian Klopsch, Anna Skorska, Marion Ludwig, Ralf Gaebel, Heiko Lemcke, Gabriela Kleiner, Martin Beyer, Brigitte Vollmar, Robert David, Gustav Steinhoff
BACKGROUND/PURPOSE: Cardiac mesenchymal stem cells (MSCs) could stimulate cell-specific regenerative mechanisms after myocardial infarction (MI) depending on spatial origin, distribution, and niche regulation. We aimed at identifying and isolating tissue-specific cardiac MSCs that could contribute to regeneration. METHODS: Following permanent ligation of the left anterior descending coronary artery in rats (n = 16), early cardiac tissues and cardiac mononuclear cells (MNCs) were analyzed by immunohistology, confocal laser scanning microscopy, and flow cytometry, respectively...
October 26, 2017: European Surgical Research. Europäische Chirurgische Forschung. Recherches Chirurgicales Européennes
https://www.readbyqxmd.com/read/29068101/isolation-and-characterisation-of-human-mesenchymal-stem-cells-derived-from-synovial-fluid-by-magnetic-activated-cell-sorting-macs
#19
Zhaofeng Jia, Yujie Liang, Xiao Xu, Xingfu Li, Qisong Liu, Yangkan Ou, Li Duan, Weimin Zhu, Wei Lu, Jianyi Xiong, Daping Wang
Mesenchymal stem cells (MSCs) are the primary source of cells used for cell-based therapy in tissue engineering. Mesenchymal stem cells (MSCs) are found in synovial fluid, a source that could be conveniently used for cartilage tissue engineering. However, the purification and characterisation of SF-MSCs has been poorly documented in the literature. Here we outline an easy-to-perform approach for the isolation and culture of MSCs derived from human synovial fluid (hSF-MSCs). We have successfully purified hSF-MSCs using magnetic activated cell sorting (MACS) sorting using the MSC surface marker, CD90...
October 25, 2017: Cell Biology International
https://www.readbyqxmd.com/read/29067104/isolation-and-characterization-of-human-umbilical-cord-derived-endothelial-colony-forming-cells
#20
Hao Zhang, Yanling Tao, Saisai Ren, Haihui Liu, Hui Zhou, Jiangwei Hu, Yongyong Tang, Bin Zhang, Hu Chen
Endothelial colony-forming cells (ECFCs) are a population of endothelial progenitor cells (EPCs) that display robust proliferative potential and vessel-forming capability. Previous studies have demonstrated that a limited number of ECFCs may be obtained from adult bone marrow, peripheral blood and umbilical cord (UC) blood. The present study describes an effective method for isolating ECFCs from human UC. The ECFCs derived from human UC displayed the full properties of EPCs. Analysis of the growth kinetics, cell cycle and colony-forming ability of the isolated human UC-ECFCs indicated that the cells demonstrated properties of stem cells, including relative stability and rapid proliferation in vitro...
November 2017: Experimental and Therapeutic Medicine
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