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Stem Cell Research

Sophia Fernandes, Shruti Tembe, Sanjay Singh, Shakti Vardhan, Velu Nair, Vaijayanti Kale, Lalita Limaye
No abstract text is available yet for this article.
February 13, 2019: Stem Cell Research
Dominic Lenz, Christian Staufner, Selina Wächter, Maike Hagedorn, Juliane Ebersold, Gudrun Göhring, Stefan Kölker, Georg F Hoffmann, Sabine Jung-Klawitter
Fibroblasts of a patient with Infantile Liver Failure Syndrome 2 (OMIM #616483) due to a homozygous missense variant in the neuroblastoma amplified sequence gene (NBAS; c.[2708T>G]; c.[2708T>G]/p.[Leu903Arg]; p.[Leu903Arg]) were reprogrammed to iPSCs using the Cytotune®-iPS 2.0 Sendai Reprogramming Kit (Invitrogen) delivering the reprogramming factors Oct3/4, Sox2, c-Myc and Klf4. Cells showed a normal karyotype. Pluripotency of DHMCi004-A was proven using immunohistochemistry, RT-PCR analysis, flow cytometry and differentiation into all three germ layers using the STEMdiff™ Trilineage Differentiation Kit (Stemcell Technologies)...
February 11, 2019: Stem Cell Research
Alexandra Haase, Wolfgang Glienke, Lena Engels, Gudrun Göhring, Ruth Esser, Lubomir Arseniev, Ulrich Martin
The utilization of human induced pluripotent stem cells (hiPSCs) for disease modeling and drug discovery is already reality, and several first-in-man-applications as cellular therapeutics have been initiated. Implementation of good manufacturing practice (GMP)-compliant protocols for the generation of hiPSC lines is crucial to increase the application safety as well as to fulfil the legal requirements for clinical trials approval. Here we describe the development of a GMP-compatible protocol for the reprogramming of CD34+ hematopoietic stem cells from peripheral blood (CD34+ PBHSC) into hiPSCs using Sendai virus-based reprogramming vectors...
February 11, 2019: Stem Cell Research
Marco Bardenbacher, Barbara Ruder, Nathalie Britzen-Laurent, Benjamin Schmid, Maximilian Waldner, Elisabeth Naschberger, Michael Scharl, Werner Müller, Claudia Günther, Christoph Becker, Michael Stürzl, Philipp Tripal
The aberrant regulation of the epithelial barrier integrity is involved in many diseases of the digestive tract, including inflammatory bowel diseases and colorectal cancer. Intestinal epithelial cell organoid cultures provide new perspectives for analyses of the intestinal barrier in vitro. However, established methods of barrier function analyses from two dimensional cultures have to be adjusted to the analysis of three dimensional organoid structures. Here we describe the methodology for analysis of epithelial barrier function and molecular regulation in intestinal organoids...
February 7, 2019: Stem Cell Research
Zuzana Kahounová, Eva Slabáková, Lucia Binó, Ján Remšík, Radek Fedr, Jan Bouchal, Radek Vrtěl, Lucie Jurečková, Volodymyr Porokh, Darja Páralová, Aleš Hampl, Karel Souček
Human induced pluripotent stem cell line was generated from commercially available primary human prostate fibroblasts HPrF derived from a fetus, aged 18-24 weeks of gestation. The fibroblast cell line was reprogrammed with Yamanaka factors (OCT4, SOX2, c-MYC, KLF4) using CytoTune™-iPS 2.0 Sendai Reprogramming Kit. Pluripotency of the derived transgene-free iPS cell line was confirmed both in vitro by detecting the expression of factors of pluripotency on a single-cell level, and in vivo using teratoma formation assay...
February 7, 2019: Stem Cell Research
Henriette R Frederiksen, Bjørn Holst, Ulrike A Mau-Holzmann, Kristine Freude, Benjamin Schmid
Alzheimer's disease (AD) is the most common form of dementia. Mutations in the gene PSEN1 encoding Presenilin1 are known to cause familial forms of AD with early age of onset. The most common mutation in the PSEN1 gene is the E280A mutation. iPSCs are an optimal choice for modeling AD, as they can be differentiated in vitro into neural cells. Here, we report the generation of two isogenic iPSC lines with either a homozygous or a heterozygous E280A mutation in the PSEN1 gene. The mutation was introduced into an iPSC line from a healthy individual using the CRISPR-Cas9 technology...
February 7, 2019: Stem Cell Research
Mercedes Tomé, Jan Tchorz, Martin Gassmann, Bernhard Bettler
Notch signalling regulates neural stem cell (NSC) proliferation, differentiation and survival for the correct development and functioning of the central nervous system. Overactive Notch2 signalling has been associated with poor prognosis of aggressive brain tumours, such as glioblastoma multiforme (GBM). We recently reported that constitutive expression of the Notch2 intracellular domain (N2ICD) enhances proliferation and gliogenesis in NSCs. Here, we investigated the mechanism by which Notch2 promotes resistance to apoptosis of NSCs to cytotoxic insults...
February 7, 2019: Stem Cell Research
Thomas Klein, Katharina Klug, Lisa Henkel, Chee Keong Kwok, Frank Edenhofer, Eva Klopocki, Ingo Kurth, Nurcan Üçeyler
Induced pluripotent stem cells (iPSC) were derived from human dermal fibroblasts (HDF) of two siblings with small fiber neuropathy (SFN) potentially based on the same variation in SCN10A but exhibiting diverse disease phenotypes. HDF were reprogrammed using a non-integrating mRNA approach and showed robust expression of pluripotency markers. iPSC displayed no chromosomal aberrations and were differentiated into all three germ-layers. These two cell lines with a familial genetic background may provide a useful in vitro tool to investigate the underlying mechanisms leading to different phenotypes caused by the same variation...
February 2, 2019: Stem Cell Research
E Ilker Ozay, Jyothi Vijayaraghavan, Gabriela Gonzalez-Perez, Sudarvili Shanthalingam, Heather L Sherman, Daniel T Garrigan, Karthik Chandiran, Joe A Torres, Barbara A Osborne, Gregory N Tew, Igor I Slukvin, Ross A Macdonald, Kilian Kelly, Lisa M Minter
The immune-mediated tissue destruction of graft-vs-host disease (GvHD) remains a major barrier to greater use of hematopoietic stem cell transplantation (HSCT). Mesenchymal stem cells (MSCs) have intrinsic immunosuppressive qualities and are being actively investigated as a therapeutic strategy for treating GvHD. We characterized Cymerus™ MSCs, which are derived from adult, induced pluripotent stem cells (iPSCs), and show they display surface markers and tri-lineage differentiation consistent with MSCs isolated from bone marrow (BM)...
February 1, 2019: Stem Cell Research
Satoshi Matsui, Miyuki Ochiai, Katsutaro Yasuda, Shin-Ichi Mae, Maki Kotaka, Taro Toyoda, Takuya Yamamoto, Kenji Osafune
Cholangiocytes are the epithelial cells that line bile ducts, and ductal plate malformation is a developmental anomaly of bile ducts that causes severe congenital biliary disorders. However, because of a lack of specific marker genes, methods for the stepwise differentiation and isolation of human induced pluripotent stem cell (hiPSC)-derived cholangiocyte progenitors at ductal plate stages have not been established. We herein generated an AQP1-GFP reporter hiPSC line and developed a combination treatment with transforming growth factor (TGF) β2 and epidermal growth factor (EGF) to induce hiPSC-derived hepatoblasts into AQP1+ cells in vitro...
January 31, 2019: Stem Cell Research
Qinxian Zhang, Zhuo Li, Huifang Sun, Shoutao Zhang, Jin Zhang, Yanlin Wang, Hui Fang, Yuming Xu
Several SLC20A2 mutations have been implicated as potential causes of Fahr's disease, a subtype of primary familial brain calcification (PFBC), but very few patient-derived induced pluripotent stem cell (iPSC) models have been established. We have identified a novel SLC20A2 mutation in a family with Fahr's disease. We subsequently obtained dermal fibroblasts from a patient in this family. These fibroblasts were successfully transformed into iPSCs by employing episomal plasmids expressing OCT3/4, SOX2, KLF4, LIN28, and L-MYC...
January 28, 2019: Stem Cell Research
Sophia Fernandes, Shruti Tembe, Sanjay Singh, Shakti Vardhan, Velu Nair, Vaijayanti Kale, Lalita Limaye
Here we report the reprogramming of CD34+ cells obtained from UCB of a healthy donor female child belonging to the Indian ethnic population. These CD34+ cells were subjected to nucleofection for delivery of episomal vectors expressing Oct4, Sox2, L-Myc, Lin28, Klf4 and p53DD (negative mutation in p53). The iPSC colonies expressed pluripotency markers as detected by PCR, immunofluorescence and flow-cytometry. The removal of plasmid was confirmed by its absence in cells at higher passages. Karyotype analysis revealed a stable genome...
January 26, 2019: Stem Cell Research
Giovanna Piovani, Gaetana Lanzi, Rosalba Monica Ferraro, Stefania Masneri, Chiara Barisani, Giulia Savio, Silvia Clara Giliani
The Cri du Chat Syndrome (CdCS) is a genetic disease resulting from variable size deletion occurring on the short arm of chromosome 5. The main clinical features are a high-pitched monochromatic cry, microcephaly, severe psychomotor and mental retardation with characteristics of autism spectrum disorders such as hand flapping, obsessive attachments to objects, twirling objects, repetitive movements, and rocking. We reprogrammed to pluripotency peripheral blood mononuclear cells derived from a patient carrying large deletion on the short arm of chromosome 5, using a commercially available non-integrating expression system...
January 26, 2019: Stem Cell Research
Ahmad Al-Moujahed, Bo Tian, Nikolaos E Efstathiou, Eleni K Konstantinou, Mien Hoang, Haijiang Lin, Joan W Miller, Demetrios G Vavvas
The molecular mechanisms involved in induced pluripotent stem cells (iPSCs) generation are poorly understood. The cell death machinery of apoptosis-inducing caspases have been shown to facilitate the process of iPSCs reprogramming. However, the effect of other cell death processes, such as programmed necrosis (necroptosis), on iPSCs induction has not been studied. In this study, we investigated the role of receptor-interacting protein kinase 3 (RIP3), an essential regulator of necroptosis, in reprogramming mouse embryonic fibroblast cells (MEFs) into iPSCs...
January 23, 2019: Stem Cell Research
Aarne Fleischer, Iván M Lorenzo, Esther Palomino, Trond Aasen, Fernando Gómez, Miguel Servera, Víctor J Asensio, Víctor Gálvez, Juan Carlos Izpisúa-Belmonte, Daniel Bachiller
No abstract text is available yet for this article.
January 22, 2019: Stem Cell Research
Marina Riera, Achchhe Patel, Borja Corcostegui, Stanley Chang, Barbara Corneo, Janet R Sparrow, Esther Pomares
A human induced pluripotent stem cell (iPSC) line was generated from a female patient affected by autosomal recessive retinitis pigmentosa with two mutations in the USH2A gene: c.2209C > T (p.Arg737Ter) and c.8693A > C (p.Tyr2898Ser). Skin fibroblasts were infected with Sendai virus containing the Yamanaka factors and the resulting cells were fully characterized to confirm successful reprogramming. The iPSC line expressed several pluripotency markers, could generate the three germ layers, had a normal karyotype, carried the two USH2A mutations and was free of Sendai virus...
January 17, 2019: Stem Cell Research
Marina Riera, Achchhe Patel, Borja Corcostegui, Stanley Chang, Janet R Sparrow, Esther Pomares, Barbara Corneo
Retinitis pigmentosa (RP) refers to a clinical and genetic heterogeneous group of inherited retinal degenerations characterized by photoreceptor cell death. In this work, we have generated an induced pluripotent stem cell (iPSC) line derived from a RP patient with two heterozygous mutations in the cGMP-specific phosphodiesterase 6A alpha subunit (PDE6A) gene. Skin fibroblasts were generated and reprogrammed by using a Sendai virus-based approach. The iPSC line had a normal karyotype, carried the two PDE6A mutations, expressed pluripotency markers and could generate endoderm, mesoderm and ectoderm in vitro...
January 17, 2019: Stem Cell Research
K R Valetdinova, M A Maretina, M L Kuranova, E V Grigor'eva, Y M Minina, E A Kizilova, A V Kiselev, S P Medvedev, V S Baranov, S M Zakian
Spinal muscular atrophy (SMA) is a neuromuscular disease caused by deletion or mutation in SMN1 gene. SMA human induced pluripotent stem cells (iPSCs) represent a useful and valid model for the study of the disorder, as they provide in vitro the target cells. We generated iPSCs from a SMA type I patient and SMA type II patient by using non-integrating episomal plasmid vectors. The resulting iPSCs are episomal-free, express pluripotency markers, display a normal karyotype, retain the mutation (homozygous deletion of SMN1) and are able to differentiate into the three germ layers...
January 11, 2019: Stem Cell Research
Benjamin Schmid, Kennie R Prehn, Natakarn Nimsanor, Blanca Irene Aldana Garcia, Ulla Poulsen, Ida Jørring, Mikkel A Rasmussen, Christian Clausen, Ulrike A Mau-Holzmann, Sarayu Ramakrishna, Ravi Muddashetty, Rachel Steeg, Kevin Bruce, Peter Mackintosh, Andreas Ebneth, Bjørn Holst, Alfredo Cabrera-Socorro
Alzheimer's disease (AD) is the most frequent neurodegenerative disease amongst the elderly. The SNPs rs429358 and rs7412 in the APOE gene are the most common risk factor for sporadic AD, and there are three different alleles commonly referred to as APOE-ε2, APOE-ε3 and APOE-ε4. Induced pluripotent stem cells (iPSCs) hold great promise to model AD as such cells can be differentiated in vitro to the required cell type. Here we report the use of CRISPR/Cas9 technology employed on iPSCs from a healthy individual with an APOE-ε3/ε4 genotype to obtain isogenic APOE-ε2/ε2, APOE-ε3/ε3, APOE-ε4/ε4 lines as well as an APOE-knock-out line...
January 4, 2019: Stem Cell Research
L Auboyer, C Monzo, D Wallon, A Rovelet-Lecrux, A Gabelle, I Gazagne, V Cacheux, S Lehmann, C Crozet
Induced pluripotent stem cells (iPSC) were generated from skin fibroblasts obtained from a 50 year-old patient suffering from Alzheimer's disease and carrying a G217D causal mutation on presenilin 1 (PSEN1). iPSCs were obtained following reprogramming using the integration-free Sendai Virus system which allows expression of the Yamanaka factors. Verification of their pluripotency was achieved by demonstrating the expression of pluripotency markers and their differentiation potential into the three primary germ layers...
January 3, 2019: Stem Cell Research
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