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Female germ stem cell

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https://www.readbyqxmd.com/read/27789408/lymphoblast-derived-integration-free-ips-cell-line-from-a-female-67-year-old-alzheimer-s-disease-patient-with-trem2-r47h-missense-mutation
#1
Friederike Schröter, Kristel Sleegers, Elise Cuyvers, Martina Bohndorf, Wasco Wruck, Christine Van Broeckhoven, James Adjaye
Human lymphoblast cells from a female patient diagnosed with Alzheimer's disease (AD) possessing the missense mutation TREM2 p.R47H were used to generate integration-free induced pluripotent stem cells (iPSCs) employing episomal plasmids expressing OCT4, SOX2, NANOG, LIN28, c-MYC and L-MYC. The iPSCs retained the TREM2 mutation, and were defined as pluripotent based on (i) expression of pluripotent-associated markers, (ii) embryoid body-based differentiation into cell types representative of the three germ layers and (iii) the similarity between the transcriptomes of the iPSC line and the human embryonic stem cell line H1 with a Pearson correlation of 0...
October 20, 2016: Stem Cell Research
https://www.readbyqxmd.com/read/27789404/establishment-of-ehmt1-mutant-induced-pluripotent-stem-cell-ipsc-line-from-a-11-year-old-kleefstra-syndrome-ks-patient-with-autism-and-normal-intellectual-performance
#2
Eszter Varga, Csilla Nemes, Zsuzsanna Táncos, István Bock, Sára Berzsenyi, György Lévay, Viktor Román, Julianna Kobolák, András Dinnyés
Peripheral blood was collected from a clinically characterized female Kleefstra syndrome patient with a heterozygous, de novo, premature termination codon (PTC) mutation (NM_024757.4(EHMT1):c.3413G>A; p.Trp1138Ter). Peripheral blood mononuclear cells (PBMCs) were reprogrammed with the human OSKM transcription factors using the Sendai-virus (SeV) delivery system. The pluripotency of transgene-free iPSC line was verified by the expression of pluripotency-associated markers and by in vitro spontaneous differentiation towards the 3 germ layers...
October 2, 2016: Stem Cell Research
https://www.readbyqxmd.com/read/27789401/generation-of-human-induced-pluripotent-stem-cell-ipsc-line-from-an-unaffected-female-carrier-of-mucopolysaccharidosis-type-ii-mps-ii-disorder
#3
Eszter Varga, Csilla Nemes, Eszter Kovács, István Bock, Norbert Varga, Anita Fehér, András Dinnyés, Julianna Kobolák
Peripheral blood was collected from a 39-year-old unaffected female carrier of an X-linked recessive mutation of Iduronate 2-sulfatase gene (NM_000202.7(IDS):c.85C>T) causing MPS II (OMIM 309900). Peripheral blood mononuclear cells (PBMCs) were reprogrammed by lentiviral delivery of a self-silencing hOKSM polycistronic vector. The pluripotency of iPSC line was confirmed by the expression of pluripotency-associated markers and in vitro spontaneous differentiation towards the 3 germ layers. The iPSC showed normal karyotype...
October 3, 2016: Stem Cell Research
https://www.readbyqxmd.com/read/27750280/reconstitution-in-vitro-of-the-entire-cycle-of-the-mouse-female-germ-line
#4
Orie Hikabe, Nobuhiko Hamazaki, Go Nagamatsu, Yayoi Obata, Yuji Hirao, Norio Hamada, So Shimamoto, Takuya Imamura, Kinichi Nakashima, Mitinori Saitou, Katsuhiko Hayashi
The female germ line undergoes a unique sequence of differentiation processes that confers totipotency to the egg. The reconstitution of these events in vitro using pluripotent stem cells is a key achievement in reproductive biology and regenerative medicine. Here we report successful reconstitution in vitro of the entire process of oogenesis from mouse pluripotent stem cells. Fully potent mature oocytes were generated in culture from embryonic stem cells and from induced pluripotent stem cells derived from both embryonic fibroblasts and adult tail tip fibroblasts...
October 17, 2016: Nature
https://www.readbyqxmd.com/read/27734353/single-cell-lineage-analysis-of-oogenesis-in-mice
#5
Lei Lei, Allan C Spradling
Lineage analysis is widely used because it provides a very powerful tool for characterizing the developmental behavior of the cells in vivo. In this chapter, we describe a particularly informative variant of lineage analysis that we term "single-cell lineage analysis". As in traditional lineage analysis, the method employs a Tamoxifen (Tmx)-inducible CAGCreER mouse line, which is crossed to an R26R reporter line that can be activated by Cre-mediated DNA recombination. However, instead of driving CreER at a high level within a subset of cells defined by a particular promoter, CreER is driven with a generic promoter that is active in essentially all cells throughout the lifespan of the mouse...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27734351/identification-of-germ-line-stem-cells-in-zebrafish
#6
Bruce W Draper
Both male and female zebrafish have a population of germ-line stem cells that produce gametes throughout the life of the fish. These cells localize to specific regions in the gonads and can be identified because they uniquely express the nanos2 gene, which encodes a conserved regulator of translation. A method is presented here for identifying germ-line stem cells in the ovary and testis using a combined protocol for whole-mount fluorescent RNA in situ hybridization to detect nanos2 mRNA and immunofluorescence to detect the pan-germ cell marker Vasa...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27734350/measurement-of-mrna-poly-a-tail-lengths-in-drosophila-female-germ-cells-and-germ-line-stem-cells
#7
Aymeric Chartier, Willy Joly, Martine Simonelig
mRNA regulation by poly(A) tail length variations plays an important role in many developmental processes. Recent advances have shown that, in particular, deadenylation (the shortening of mRNA poly(A) tails) is essential for germ-line stem cell biology in the Drosophila ovary. Therefore, a rapid and accurate method to analyze poly(A) tail lengths of specific mRNAs in this tissue is valuable. Several methods of poly(A) test (PAT) assays have been reported to measure mRNA poly(A) tail lengths in vivo. Here, we describe two of these methods (PAT and ePAT) that we have adapted for Drosophila ovarian germ cells and germ-line stem cells...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27699806/germline-stem-cells-are-critical-for-sexual-fate-decision-of-germ-cells
#8
Minoru Tanaka
Egg or sperm? The mechanism of sexual fate decision in germ cells has been a long-standing issue in biology. A recent analysis identified foxl3 as a gene that determines the sexual fate decision of germ cells in the teleost fish, medaka. foxl3/Foxl3 acts in female germline stem cells to repress commitment into male fate (spermatogenesis), indicating that the presence of mitotic germ cells in the female is critical for continuous sexual fate decision of germ cells in medaka gonads. Interestingly, foxl3 is found in most vertebrate genomes except for mammals...
December 2016: BioEssays: News and Reviews in Molecular, Cellular and Developmental Biology
https://www.readbyqxmd.com/read/27659063/an-efficient-method-for-generating-a-germ-cell-depleted-animal-model-for-studies-related-to-spermatogonial-stem-cell-transplantation
#9
Nirmalya Ganguli, Neerja Wadhwa, Abul Usmani, Neetu Kunj, Nilanjana Ganguli, Rajesh Kumar Sarkar, Soma M Ghorai, Subeer S Majumdar
BACKGROUND: Spermatogonial stem cell (SSC) transplantation (SSCT) has become important for conservation of endangered species, transgenesis and for rejuvenating testes which have lost germ cells (Gc) due to gonadotoxic chemotherapy or radiotherapy during the prepubertal phase of life. Creating a germ cell-depleted animal model for transplantation of normal or gene-transfected SSC is a prerequisite for such experimental studies. Traditionally used intraperitoneal injections of busulfan to achieve this are associated with painful hematopoietic toxicity and affects the wellbeing of the animals...
September 22, 2016: Stem Cell Research & Therapy
https://www.readbyqxmd.com/read/27642137/global-landscape-and-regulatory-principles-of-dna-methylation-reprogramming-for-germ-cell-specification-by-mouse-pluripotent-stem-cells
#10
Kenjiro Shirane, Kazuki Kurimoto, Yukihiro Yabuta, Masashi Yamaji, Junko Satoh, Shinji Ito, Akira Watanabe, Katsuhiko Hayashi, Mitinori Saitou, Hiroyuki Sasaki
Specification of primordial germ cells (PGCs) activates epigenetic reprogramming for totipotency, the elucidation of which remains a fundamental challenge. Here, we uncover regulatory principles for DNA methylation reprogramming during in vitro PGC specification, in which mouse embryonic stem cells (ESCs) are induced into epiblast-like cells (EpiLCs) and then PGC-like cells (PGCLCs). While ESCs reorganize their methylome to form EpiLCs, PGCLCs essentially dilute the EpiLC methylome at constant, yet different, rates between unique sequence regions and repeats...
October 10, 2016: Developmental Cell
https://www.readbyqxmd.com/read/27572922/c-terminal-residues-specific-to-vasa-among-dead-box-helicases-are-required-for-its-functions-in-pirna-biogenesis-and-embryonic-patterning
#11
Mehrnoush Dehghani, Paul Lasko
The DEAD-box RNA helicase Vasa (Vas, also known as DDX4) is required for germ cell development. In Drosophila, analysis of hypomorphic mutations has implicated maternally expressed Vas in germ cell formation and posterior embryonic patterning. vas-null females, which rarely complete oogenesis, exhibit defects in mitotic progression of germline stem cells, Piwi-interacting RNA (piRNA)-mediated transposon silencing, and translation of Gurken (Grk), an EGFR ligand. The carboxy-terminal region of Vas orthologs throughout the animal kingdom consists of several acidic residues as well as an invariant tryptophan in the penultimate or ultimate position (Trp660 in Drosophila melanogaster)...
August 29, 2016: Development Genes and Evolution
https://www.readbyqxmd.com/read/27558602/derivation-of-the-clinical-grade-human-embryonic-stem-cell-line-rce015-a-rc-11
#12
P A De Sousa, B J Tye, K Bruce, P Dand, G Russell, D M Collins, A Greenshields, K McDonald, H Bradburn, A Laurie, M Canham, T Kunath, J M Downie, M Bateman, A Courtney
The human embryonic stem cell line RCe015-A (RC-11) was derived under quality assured compliance with UK regulation, European Union Directives and International guidance for tissue procurement, processing and storage according to Good Manufacturing Practice (GMP) standards. The cell line was derived from a fragmented cleavage stage embryo voluntarily donated as unsuitable or surplus to fertility requirements following informed consent. RCe015-A (RC-11) shows normal pluripotency marker expression and differentiation to the three germ layers in vitro and in vivo...
May 7, 2016: Stem Cell Research
https://www.readbyqxmd.com/read/27558600/integration-free-t-cell-derived-human-induced-pluripotent-stem-cells-ipscs-from-a-patient-with-recessive-dystrophic-epidermolysis-bullosa-rdeb-carrying-two-compound-heterozygous-mutations-in-the-col7a1-gene
#13
Munenari Itoh, Shiho Kawagoe, Katsuto Tamai, Hirotaka James Okano, Hidemi Nakagawa
Expanded human T cells from a Japanese female with recessive dystrophic epidermolysis bullosa (RDBE) were used to generate integration-free induced pluripotent stem cells (iPSCs) by exogenous expression of four reprogramming factors, OCT3/4, SOX2, cMYC, KLF4, using Sendai virus vector (SeVdp). The authenticity of established iPSC line, RDEB-iPSC26, was confirmed by the expressions of stem cell markers and the differentiation capability into three germ layer. RDEB-iPSC26 may be a useful cell resource for the establishment of in vitro RDEB modeling and the study for developing gene and cell therapy...
May 17, 2016: Stem Cell Research
https://www.readbyqxmd.com/read/27558596/derivation-of-the-clinical-grade-human-embryonic-stem-cell-line-rce021-a-rc-17
#14
P A De Sousa, B J Tye, K Bruce, P Dand, G Russell, D M Collins, A Greenshields, K McDonald, H Bradburn, D Allan, A Laurie, T Kunath, M A Canham, J M Downie, M Bateman, A Courtney
The human embryonic stem cell line RCe021-A (RC-17) was derived under quality assured compliance with UK regulation, European Union Directives and International guidance for tissue procurement, processing and storage according to Good Manufacturing Practice (GMP) standards. The cell line was derived from a day 3 embryo voluntarily donated as unsuitable or surplus to fertility requirements following informed consent. RCe021-A (RC-17) shows normal pluripotency marker expression and differentiation to the three germ layers in vitro...
April 30, 2016: Stem Cell Research
https://www.readbyqxmd.com/read/27557587/isolation-of-mammalian-oogonial-stem-cells-by-antibody-based-fluorescence-activated-cell-sorting
#15
Deanna M Navaroli, Jonathan L Tilly, Dori C Woods
The ability to isolate and subsequently culture mitotically active female germ cells from adult ovaries, referred to as either oogonial stem cells (OSCs) or adult female germline stem cells (aFGSCs), has provided a robust system to study female germ cell development under multiple experimental conditions, and in many species. Flow cytometry or fluorescence-activated cell sorting (FACS) is an integral part of many isolation and characterization protocols. Here, we provide methodological details for antibody-based flow cytometric isolation of OSCs using antibodies specific for external epitopes of the proteins Ddx4 or Ifitm3, alone or in combination with the use of fluorescent reporter mice...
2016: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27498563/wolbachia-protein-tomo-targets-nanos-mrna-and-restores-germ-stem-cells-in-drosophila-sex-lethal-mutants
#16
Manabu Ote, Morio Ueyama, Daisuke Yamamoto
Wolbachia, endosymbiotic bacteria prevalent in invertebrates, manipulate their hosts in a variety of ways: they induce cytoplasmic incompatibility, male lethality, male-to-female transformation, and parthenogenesis. However, little is known about the molecular basis for host manipulation by these bacteria. In Drosophila melanogaster, Wolbachia infection makes otherwise sterile Sex-lethal (Sxl) mutant females capable of producing mature eggs. Through a functional genomic screen for Wolbachia genes with growth-inhibitory effects when expressed in cultured Drosophila cells, we identified the gene WD1278 encoding a novel protein we call toxic manipulator of oogenesis (TomO), which phenocopies some of the Wolbachia effects in Sxl mutant D...
September 12, 2016: Current Biology: CB
https://www.readbyqxmd.com/read/27473657/mouse-d1pas1-a-dead-box-rna-helicase-is-required-for-the-completion-of-first-meiotic-prophase-in-male-germ-cells
#17
Hiroki Inoue, Narumi Ogonuki, Michiko Hirose, Yuki Hatanaka, Shogo Matoba, Shinichiro Chuma, Kimio Kobayashi, Shigeharu Wakana, Junko Noguchi, Kimiko Inoue, Kentaro Tanemura, Atsuo Ogura
D1Pas1 is a mouse autosomal DEAD-box RNA helicase expressed predominantly in the testis. To assess its possible function, we generated D1Pas1-deficient mice using embryonic stem cells with a targeted D1Pas1 allele. Deletion of D1Pas1 did not cause noticeable embryonic defects or death, indicating that D1Pas1 is not essential for embryogenesis. Whereas homozygous knockout female mice showed normal reproductive performance, homozygous knockout male mice were completely sterile. The seminiferous epithelium of D1Pas1-deficient males contained no spermatids or spermatozoa because of spermatogenic arrest at the late pachytene stage...
September 16, 2016: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/27410401/a-structurally-and-functionally-common-unit-in-testes-and-ovaries-of-medaka-oryzias-latipes-a-teleost-fish
#18
Toshiya Nishimura, Shuhei Nakamura, Minoru Tanaka
Testes and ovaries are structurally and functionally different organs, but they originate from the same gonadal primordium. Here, we propose how a tissue composed of germ cells and sox9b-expressing cells underlain by a basement membrane develops a functionally common unit that reserves sexually indifferent or unfixed germline stem cells in both the ovaries and testes of adult medaka. During testicular development, the unit expands and gives rise to lobules where germline stem cells continuously produce a tremendous amount of sperm, while the unit in the female becomes germinal cradles that represent niche areas for neo-oogenesis...
2016: Sexual Development: Genetics, Molecular Biology, Evolution, Endocrinology, Embryology, and Pathology of Sex Determination and Differentiation
https://www.readbyqxmd.com/read/27383240/perchlorate-exposure-reduces-primordial-germ-cell-number-in-female-threespine-stickleback
#19
Ann M Petersen, Nathanial C Earp, Mandy E Redmond, John H Postlethwait, Frank A von Hippel, C Loren Buck, William A Cresko
Perchlorate is a common aquatic contaminant that has long been known to affect thyroid function in vertebrates, including humans. More recently perchlorate has been shown to affect primordial sexual differentiation in the aquatic model fishes zebrafish and threespine stickleback, but the mechanism has been unclear. Stickleback exposed to perchlorate from fertilization have increased androgen levels in the embryo and disrupted reproductive morphologies as adults, suggesting that perchlorate could disrupt the earliest stages of primordial sexual differentiation when primordial germ cells (PGCs) begin to form the gonad...
2016: PloS One
https://www.readbyqxmd.com/read/27364467/the-wnt-pathway-limits-bmp-signaling-outside-of-the-germline-stem-cell-niche-in-drosophila-ovaries
#20
Violaine I Mottier-Pavie, Victor Palacios, Susan Eliazer, Shane Scoggin, Michael Buszczak
The mechanisms that modulate and limit the signaling output of adult stem cell niches remain poorly understood. To gain further insights into how these microenvironments are regulated in vivo, we performed a candidate gene screen designed to identify factors that restrict BMP signal production to the cap cells that comprise the germline stem cell (GSC) niche of Drosophila ovaries. Through these efforts, we found that disruption of Wnt4 and components of the canonical Wnt pathway results in a complex germ cell phenotype marked by an expansion of GSC-like cells, pre-cystoblasts and cystoblasts in young females...
September 1, 2016: Developmental Biology
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