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https://www.readbyqxmd.com/read/29786554/a-human-ipsc-line-capable-of-differentiating-into-functional-macrophages-expressing-zsgreen-a-tool-for-the-study-and-in-vivo-tracking-of-therapeutic-cells
#1
Martha Lopez-Yrigoyen, Antonella Fidanza, Luca Cassetta, Richard A Axton, A Helen Taylor, Jose Meseguer-Ripolles, Anestis Tsakiridis, Valerie Wilson, David C Hay, Jeffrey W Pollard, Lesley M Forrester
We describe the production of a human induced pluripotent stem cell (iPSC) line, SFCi55-ZsGr, that has been engineered to express the fluorescent reporter gene, ZsGreen, in a constitutive manner. The CAG-driven ZsGreen expression cassette was inserted into the AAVS1 locus and a high level of expression was observed in undifferentiated iPSCs and in cell lineages derived from all three germ layers including haematopoietic cells, hepatocytes and neurons. We demonstrate efficient production of terminally differentiated macrophages from the SFCi55-ZsGreen iPSC line and show that they are indistinguishable from those generated from their parental SFCi55 iPSC line in terms of gene expression, cell surface marker expression and phagocytic activity...
July 5, 2018: Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences
https://www.readbyqxmd.com/read/29782016/in-vivo-gene-transfer-to-the-rabbit-common-carotid-artery-endothelium
#2
Bradley K Wacker, Lianxiang Bi, David A Dichek
The goal of this method is to introduce a transgene into the endothelium of isolated segments of both rabbit common carotid arteries. The method achieves focal endothelial-selective transgenesis, thereby allowing an investigator to determine the biological roles of endothelial-expressed transgenes and to quantify the in vivo transcriptional activity of DNA sequences in large artery endothelial cells. The method uses surgical isolation of rabbit common carotid arteries and an arteriotomy to deliver a transgene-expressing viral vector into the arterial lumen...
May 6, 2018: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29778975/establishment-of-an-integration-free-induced-pluripotent-stem-cell-line-musii005-a-from-exfoliated-renal-epithelial-cells
#3
Bootsakorn Boonkaew, Weeradee Thummavichit, Ratchapong Netsrithong, Chinnavuth Vatanashevanopakorn, Kovit Pattanapanyasat, Methichit Wattanapanitch
Human induced pluripotent stem cells (iPSCs) were generated from exfoliated renal epithelial cells isolated from a urine sample of a 31-year-old healthy woman. Epithelial cells were characterized for the expression of E-cadherin and reprogrammed using non-integrating Sendai viral vectors. The urine-derived iPSC line (designated as MUSIi005-A) was karyotypically normal, expressed pluripotent markers, differentiated into cells of three embryonic germ layers, and showed no viral and transgene expressions at passage 29...
May 16, 2018: Stem Cell Research
https://www.readbyqxmd.com/read/29769727/microbial-signals-drive-pre-leukaemic-myeloproliferation-in-a-tet2-deficient-host
#4
Marlies Meisel, Reinhard Hinterleitner, Alain Pacis, Li Chen, Zachary M Earley, Toufic Mayassi, Joseph F Pierre, Jordan D Ernest, Heather J Galipeau, Nikolaus Thuille, Romain Bouziat, Manuel Buscarlet, Daina L Ringus, Yitang Wang, Ye Li, Vu Dinh, Sangman M Kim, Benjamin D McDonald, Matthew A Zurenski, Mark W Musch, Glaucia C Furtado, Sergio A Lira, Gottfried Baier, Eugene B Chang, A Murat Eren, Christopher R Weber, Lambert Busque, Lucy A Godley, Elena F Verdú, Luis B Barreiro, Bana Jabri
Somatic mutations in tet methylcytosine dioxygenase 2 (TET2), which encodes an epigenetic modifier enzyme, drive the development of haematopoietic malignancies1-7 . In both humans and mice, TET2 deficiency leads to increased self-renewal of haematopoietic stem cells with a net developmental bias towards the myeloid lineage1,4,8,9 . However, pre-leukaemic myeloproliferation (PMP) occurs in only a fraction of Tet2-/- mice8,9 and humans with TET2 mutations1,3,5-7 , suggesting that extrinsic non-cell-autonomous factors are required for disease onset...
May 16, 2018: Nature
https://www.readbyqxmd.com/read/29761384/a-simple-multistep-protocol-for-differentiating-human-induced-pluripotent-stem-cells-into-functional-macrophages
#5
Chandrayana Mukherjee, Christine Hale, Subhankar Mukhopadhyay
Macrophages differentiated from human induced pluripotent stem cells (hiPSCs) provide an alternative new tool overcoming some of the limitations of existing models for human macrophages, such as human macrophage-like cell lines and primary monocyte-derived macrophages. A combination of different cytokines and growth factors can differentiate hiPSCs toward myeloid lineage. Here we describe a simple multistep protocol for differentiating hiPSCs into functional macrophages. This includes derivation of three germ-line containing embryoid bodies (EBs) from iPSCs, generation of myeloid precursors from EBs, and finally maturation of myeloid precursors into functional macrophages...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29753274/blood-derived-integration-free-ips-cell-line-ukbi011-a-from-a-diagnosed-male-alzheimer-s-disease-patient-with-apoe-%C3%A9-4-%C3%A9-4-genotype
#6
Michael Peitz, Tamara Bechler, Catrin Cornelia Thiele, Monika Veltel, Melanie Bloschies, Klaus Fliessbach, Alfredo Ramirez, Oliver Brüstle
Alzheimer's disease (AD) is most the frequent neurodegenerative disease, and the APOE ε4 allele is the most prominent risk factor for late-onset AD. Here, we present an iPSC line generated from peripheral blood cells of a male AD patient employing Sendai virus vectors encoding the transcription factors OCT4, SOX2, KLF4 and c-MYC. The characterized iPSC line expresses typical human pluripotency markers and shows differentiation into all three germ layers, complete reprogramming vector clearance, a normal SNP genotype and maintenance of the APOE ε4/ε4 allele...
April 23, 2018: Stem Cell Research
https://www.readbyqxmd.com/read/29743854/microsatellite-instability-in-colorectal-cancer
#7
REVIEW
Jafar Nouri Nojadeh, Shahin Behrouz Sharif, Ebrahim Sakhinia
Colorectal cancer (CRC) is a heterogeneous disease that is caused by the interaction of genetic and environmental factors. Although it is one of the most common cancers worldwide, CRC would be one of the most curable cancers if it is detected in the early stages. Molecular changes that occur in colorectal cancer may be categorized into three main groups: 1) Chromosomal Instability (CIN), 2) Microsatellite Instability (MSI), and 3) CpG Island Methylator phenotype (CIMP). Microsatellites, also known as Short Tandem Repeats (STRs) are small (1-6 base pairs) repeating stretches of DNA scattered throughout the entire genome and account for approximately 3 % of the human genome...
2018: EXCLI journal
https://www.readbyqxmd.com/read/29732171/pohad-why-we-should-study-future-fathers
#8
Adelheid Soubry
The growing field of 'Developmental Origin of Health and Disease' (DOHaD) generally reflects environmental influences from mother to child. The importance of maternal lifestyle, diet and other environmental exposures before and during gestation period is well recognized. However, few epidemiological designs explore potential influences from the paternal environment on offspring health. This is surprising given that numerous animal models have provided evidence that the paternal environment plays a role in a non-genetic inheritance of pre-conceptional exposures through the male germ line...
April 2018: Environmental Epigenetics
https://www.readbyqxmd.com/read/29730572/establishment-of-a-human-embryonic-stem-cell-line-with-homozygous-tp53-r248w-mutant-by-talen-mediated-gene-editing
#9
An Xu, Ruoji Zhou, Jian Tu, Zijun Huo, Dandan Zhu, Donghui Wang, Julian A Gingold, Helen Mata, Pulivarthi H Rao, Mo Liu, Alaa M T Mohamed, Celine Shuet Lin Kong, Brittany E Jewell, Weiya Xia, Ruiying Zhao, Mien-Chie Hung, Dung-Fang Lee
Genetic mutations in TP53 contribute to multiple human cancers. Here we report the generation of a H1-p53(R248W/R248W) human embryonic stem cell line harboring a homozygous TP53 R248W mutation created by TALEN-mediated precise gene editing. The H1-p53(R248W/R248W) cell line maintains a normal karyotype, robust pluripotency gene expression, and the potential to differentiate to the three germ layers.
April 27, 2018: Stem Cell Research
https://www.readbyqxmd.com/read/29730569/derivation-and-characterization-of-the-nih-registry-human-stem-cell-line-nyscf101-under-defined-feeder-free-conditions
#10
Ana Sevilla, Eliana Forero, Matthew Zimmer, Hector Martinez, Katie Reggio, Daniel Paull, Dieter Egli, Scott Noggle
The human embryonic stem cell line NYSCFe002-A was derived from a day 6 blastocyst in feeder-free and antibiotic free conditions. The blastocyst was voluntarily donated for research as surplus after in vitro fertilization treatment following informed consent. The NYSCFe002-A line expresses all the pluripotency markers and has the potential to differentiate into all three germ layers in vitro. The line presents normal karyotype and is mycoplasma free. This line is registered as NYSCF101 on the NIH Registry.
April 30, 2018: Stem Cell Research
https://www.readbyqxmd.com/read/29692670/recent-advances-of-in-vitro-culture-systems-for-spermatogonial-stem-cells-in-mammals
#11
REVIEW
Mahesh G Sahare, Suyatno, Hiroshi Imai
Background: Spermatogonial stem cells (SSCs) in the mammalian testis are unipotent stem cells for spermatozoa. They show unique cell characteristics as stem cells and germ cells after being isolated from the testis and cultured in vitro. This review introduces recent progress in the development of culture systems for the establishment of SSC lines in mammalian species, including humans. Methods: Based on the published reports, the isolation and purification of SSCs, identification and characteristics of SSCs, and culture system for mice, humans, and domestic animals have been summarized...
April 2018: Reproductive Medicine and Biology
https://www.readbyqxmd.com/read/29679845/establishment-of-stub1-chip-mutant-induced-pluripotent-stem-cells-ipscs-from-a-patient-with-gordon-holmes-syndrome-scar16
#12
Stefanie Schuster, Yvonne Schelling, Matthis Synofzik, Philip Höflinger, Ludger Schöls, Stefan Hauser
STUB1/CHIP is a central component of cellular protein homeostasis and interacts with key proteins involved in the pathogenesis of many neurodegenerative diseases. Here, we reprogrammed human skin fibroblasts from a 12-year-old male patient with recessive spinocerebellar ataxia type 16 (OMIM #615768), carrying compound heterozygous mutations (c.355C>T, c.880A>T) in STUB1. Genomic integrity of the iPSC line HIHCNi001-A without transgene integration and genomic aberration but with maintained disease-relevant mutations was proven by SNP array analysis and Sanger sequencing while pluripotency was verified by the expression of important pluripotency markers and the capacity to differentiate into cells of all three germ layers...
April 9, 2018: Stem Cell Research
https://www.readbyqxmd.com/read/29670214/restoring-functional-neurofibromin-by-protein-transduction
#13
K Mellert, S Lechner, M Lüdeke, M Lamla, P Möller, R Kemkemer, K Scheffzek, D Kaufmann
In Neurofibromatosis 1 (NF1) germ line loss of function mutations result in reduction of cellular neurofibromin content (NF1+/-, NF1 haploinsufficiency). The Ras-GAP neurofibromin is a very large cytoplasmic protein (2818 AA, 319 kDa) involved in the RAS-MAPK pathway. Aside from regulation of proliferation, it is involved in mechanosensoric of cells. We investigated neurofibromin replacement in cultured human fibroblasts showing reduced amount of neurofibromin. Full length neurofibromin was produced recombinantly in insect cells and purified...
April 18, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29666451/exploring-the-extent-and-scope-of-epigenetic-inheritance
#14
REVIEW
Elizabeth J Radford
Environmental factors, particularly during early life, are important for the later metabolic health of the individual. In our obesogenic environment, it is of major socio-economic importance to investigate the mechanisms that contribute to the risk of metabolic ill health. Increasing evidence from a variety of model organisms suggests that non-genetically determined phenotypes, including metabolic effects such as glucose intolerance and obesity, can be passed between generations, which encourages us to revisit heredity...
April 17, 2018: Nature Reviews. Endocrinology
https://www.readbyqxmd.com/read/29658992/can-mutation-mediated-effects-occurring-early-in-development-cause-long-term-seizure-susceptibility-in-genetic-generalized-epilepsies
#15
Christopher Alan Reid, Ben Rollo, Steven Petrou, Samuel F Berkovic
Epilepsy has a strong genetic component, with an ever-increasing number of disease-causing genes being discovered. Most epilepsy-causing mutations are germ line and thus present from conception. These mutations are therefore well positioned to have a deleterious impact during early development. Here we review studies that investigate the role of genetic lesions within the early developmental window, specifically focusing on genetic generalized epilepsy (GGE). Literature on the potential pathogenic role of sub-mesoscopic structural changes in GGE is also reviewed...
May 2018: Epilepsia
https://www.readbyqxmd.com/read/29648597/secondary-bile-acids-inhibit-candida-albicans-growth-and-morphogenesis
#16
Jack Guinan, Pedro Villa, Shankar Thangamani
Candida albicans is one of the most common causes of fungal infections in humans with a significant mortality rate. However, the factors involved in C. albicans gastrointestinal (GI) colonization remain unclear. We hypothesize that secondary bile acids have direct antifungal activity against C. albicans and may play a critical role in maintaining GI colonization resistance against C. albicans. In this study, we investigated the effect of secondary bile acids including lithocholic acid (LCA) and deoxycholic acid (DCA) on C...
April 10, 2018: Pathogens and Disease
https://www.readbyqxmd.com/read/29620155/differential-expression-of-oct3-4-in-human-breast-cancer-and-normal-tissues
#17
Feng-Qi Zhao, Yogi Misra, Da-Biao Li, Marilyn P Wadsworth, David Krag, Donald Weaver, Joseph Tessitore, Da-Wei Li, Guo Zhang, Qing Tian, Katie Buss
Oct3/4, a transcription factor specifically expressed in mammalian totipotent embryonic stem and germ cells, has a critical role in the regulation and maintenance of pluripotency and self-renewal. However, reactivation of Oct3/4 expression is observed in several human breast cancer cell lines, but not in non‑malignant cells. To examine Oct3/4 expression in human primary breast carcinomas and normal breast tissues, we obtained breast tumor tissues from 28 patients and normal breast tissues from 9 women. According to quantitative polymerase chain reaction, all of the tumor tissues, irrespective of tumor type or clinicopathological status, expressed Oct3/4 mRNA at 10- to 100- fold higher levels than that in the normal breast tissues...
March 29, 2018: International Journal of Oncology
https://www.readbyqxmd.com/read/29615558/requirement-of-the-dynein-adaptor-spindly-for-mitotic-and-post-mitotic-functions-in-drosophila
#18
Giuliana D Clemente, Matthew R Hannaford, Hamze Beati, Katja Kapp, Jens Januschke, Eric R Griffis, Hans-Arno J Müller
Spindly was originally identified as a specific regulator of Dynein activity at the kinetochore. In early prometaphase, Spindly recruits the Dynein/Dynactin complex, promoting the establishment of stable kinetochore-microtubule interactions and progression into anaphase. While details of Spindly function in mitosis have been worked out in cultured human cells and in the C. elegans zygote, the function of Spindly within the context of an organism has not yet been addressed. Here, we present loss- and gain-of-function studies of Spindly using transgenic RNAi in Drosophila ...
March 30, 2018: Journal of Developmental Biology
https://www.readbyqxmd.com/read/29613829/novel-imprinted-single-cpg-sites-found-by-global-dna-methylation-analysis-in-human-parthenogenetic-induced-pluripotent-stem-cells
#19
Na Young Choi, Jin Seok Bang, Hye Jeong Lee, Yo Seph Park, Minseong Lee, Dahee Jeong, Kisung Ko, Dong Wook Han, Hyung-Min Chung, Gwang Jun Kim, Seung-Hyuk Shim, Han Sung Hwang, Kinarm Ko
Genomic imprinting is the process of epigenetic modification whereby genes are expressed in a parent-of-origin dependent manner; it plays an important role in normal growth and development. Parthenogenetic embryos contain only the maternal genome. Parthenogenetic embryonic stem cells could be useful for studying imprinted genes. In humans, mature cystic ovarian teratomas originate from parthenogenetic activation of oocytes; they are composed of highly differentiated mature tissues containing all three germ layers...
May 3, 2018: Epigenetics: Official Journal of the DNA Methylation Society
https://www.readbyqxmd.com/read/29611115/c-elegans-based-screen-identifies-lysosome-damaging-alkaloids-that-induce-stat3-dependent-lysosomal-cell-death
#20
Yang Li, Yu Zhang, Qiwen Gan, Meng Xu, Xiao Ding, Guihua Tang, Jingjing Liang, Kai Liu, Xuezhao Liu, Xin Wang, Lingli Guo, Zhiyang Gao, Xiaojiang Hao, Chonglin Yang
Lysosomes are degradation and signaling centers within the cell, and their dysfunction impairs a wide variety of cellular processes. To understand the cellular effect of lysosome damage, we screened natural small-molecule compounds that induce lysosomal abnormality using Caenorhabditis elegans (C. elegans) as a model system. A group of vobasinyl-ibogan type bisindole alkaloids (ervachinines A-D) were identified that caused lysosome enlargement in C. elegans macrophage-like cells. Intriguingly, these compounds triggered cell death in the germ line independently of the canonical apoptosis pathway...
April 2, 2018: Protein & Cell
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