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Induced pluripotent stem

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https://www.readbyqxmd.com/read/29907164/optimizing-the-method-for-generation-of-integration-free-induced-pluripotent-stem-cells-from-human-peripheral-blood
#1
Haihui Gu, Xia Huang, Jing Xu, Lili Song, Shuping Liu, Xiao-Bing Zhang, Weiping Yuan, Yanxin Li
BACKGROUND: Generation of induced pluripotent stem cells (iPSCs) from human peripheral blood provides a convenient and low-invasive way to obtain patient-specific iPSCs. The episomal vector is one of the best approaches for reprogramming somatic cells to pluripotent status because of its simplicity and affordability. However, the efficiency of episomal vector reprogramming of adult peripheral blood cells is relatively low compared with cord blood and bone marrow cells. METHODS: In the present study, integration-free human iPSCs derived from peripheral blood were established via episomal technology...
June 15, 2018: Stem Cell Research & Therapy
https://www.readbyqxmd.com/read/29906669/generation-of-gene-corrected-ipsc-line-from-parkinson-s-disease-patient-ipsc-line-with-alpha-snca-a53t-mutation
#2
Seo-Young Lee, SangKyun Jeong, Janghwan Kim, Sun-Ku Chung
Parkinson's disease (PD) is the second most common age-related neurodegenerative disorder. PD can result from a mutation of alpha-synuclein (α-SNCA), such as α-SNCA A53T. Using episomal vectors, induced pluripotent stem cells (iPSCs) were generated from skin fibroblasts with the α-SNCA A53T mutation. A huge bacterial artificial chromosome (BAC) harboring the normal α-SNCA gene successfully corrected the α-SNCA A53T-mutant iPSCs. Melting curve analysis for allelic composition indicated that the BAC DNA was precisely targeted to the α-SNCA A53T mutation allele, without random integration...
June 9, 2018: Stem Cell Research
https://www.readbyqxmd.com/read/29905375/sox2-mediated-upregulation-of-cd24-promotes-adaptive-resistance-towards-targeted-therapy-in-melanoma
#3
Laura Hüser, Sachindra, Karol Granados, Aniello Federico, Lionel Larribère, Daniel Novak, Viktor Umansky, Peter Altevogt, Jochen Utikal
Melanoma is often characterized by a constitutively active RAS-RAF-MEK-ERK pathway. For targeted therapy, BRAF inhibitors are available that are powerful in the beginning but resistance occurs rather fast. A better understanding of the mechanisms of resistance is urgently needed to increase the success of the treatment. Here, we observed that SOX2 and CD24 are upregulated upon BRAF inhibitor treatment. A similar upregulation was seen in targeted therapy-resistant, melanoma-derived induced pluripotent cancer cells (iPCCs)...
June 15, 2018: International Journal of Cancer. Journal International du Cancer
https://www.readbyqxmd.com/read/29904151/lncrna-expression-profiling-of-bmscs-in-osteonecrosis-of-the-femoral-head-associated-with-increased-adipogenic-and-decreased-osteogenic-differentiation
#4
Qingyu Wang, Qiwei Yang, Gaoyang Chen, Zhenwu Du, Ming Ren, Ao Wang, Haiyue Zhao, Zhaoyan Li, Guizhen Zhang, Yang Song
Long noncoding RNAs (lncRNAs) are critical gene expression regulators and are involved in several bone diseases. To explore the potential roles of lncRNAs in osteonecrosis of the femoral head (ONFH), we investigated for the first time the lncRNA expression profile of bone marrow mesenchymal stem cells (BMSCs) from patients with steroid-induced ONFH (SONFH) with microarray and bioinformatics analysis. A total of 1878 lncRNAs and 838 mRNAs were significantly up-regulated while 1842 lncRNAs and 1937 mRNAs were statistically down-regulated in the SONFH group compared with control group...
June 14, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29904107/pathological-modelling-of-pigmentation-disorders-associated-with-hutchinson-gilford-progeria-syndrome-hgps-revealed-an-impaired-melanogenesis-pathway-in-ips-derived-melanocytes
#5
Alessandra Lo Cicero, Manoubia Saidani, Jennifer Allouche, Anne Laure Egesipe, Lucile Hoch, Celine Bruge, Sabine Sigaudy, Annachiara De Sandre-Giovannoli, Nicolas Levy, Christine Baldeschi, Xavier Nissan
Hutchinson-Gilford Progeria Syndrome (HGPS) is a rare genetic disorder that leads to premature aging. In this study, we used induced pluripotent stem cells to investigate the hypopigmentation phenotypes observed in patients with progeria. Accordingly, two iPS cell lines were derived from cells from HGPS patients and differentiated into melanocytes. Measurements of melanin content revealed a lower synthesis of melanin in HGPS melanocytes as compared to non-pathologic cells. Analysis of the melanosome maturation process by electron microscopy revealed a lower percentage of mature, fully pigmented melanosomes...
June 14, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29902946/silver-nanoparticles-inhibit-neural-induction-in-human-induced-pluripotent-stem-cells
#6
Shigeru Yamada, Daiju Yamazaki, Yasunari Kanda
Silver nanoparticles (AgNPs) have been widely used as consumer products due to their antibacterial activities. Despite their extensive use, AgNPs have been reported to cause various types of cytotoxicity, including neurotoxicity. However, the potential action of AgNPs on early fetal development has not been elucidated. This study determined the effects of AgNPs on neural induction in human induced pluripotent stem cells (iPSCs), used as a model for human fetal stage development. It was observed that exposure to AgNPs reduced the expression of several neural differentiation marker genes, including OTX2, an early biomarker for neurogenesis in iPSCs...
June 14, 2018: Nanotoxicology
https://www.readbyqxmd.com/read/29902745/lymphoblast-derived-integration-free-ipsc-line-ad-trem2-3-from-a-74-year-old-alzheimer-s-disease-patient-expressing-the-trem2-p-r47h-variant
#7
Soraia Martins, Hatice Yigit, Martina Bohndorf, Nina Graffmann, Aurelian Robert Fiszl, Wasco Wruck, Kristel Sleegers, Christine Van Broeckhoven, James Adjaye
Human lymphoblast cells from a male diagnosed with Alzheimer's disease (AD) expressing the TREM2 p.R47H variant were used to generate integration-free induced pluripotent stem cells (iPSCs) by over-expressing episomal-based plasmids harbouring OCT4, SOX2, KLF4, LIN28, L-MYC and p53 shRNA. The derived iPSC line - AD-TREM2-3 was defined as pluripotent based on (i) expression of pluripotency-associated markers (ii) embryoid body-based differentiation into cell types representative of the three germ layers and (iii) the similarity between the transcriptome of the iPSC line and the human embryonic stem cell line H1 with a Pearson correlation of 0...
June 1, 2018: Stem Cell Research
https://www.readbyqxmd.com/read/29902661/a-feasibility-study-of-the-toxic-responses-of-human-induced-pluripotent-stem-cell-derived-hepatocytes-to-phytochemicals
#8
Tomáš Smutný, Riina Harjumäki, Liisa Kanninen, Marjo Yliperttula, Petr Pávek, Yan-Ru Lou
Herbal medicines have been increasingly used in the last three decades. Despite their popularity, safety issues with herbal products need to be addressed. We performed a feasibility study of the toxic responses of human induced pluripotent stem cell-derived hepatocytes (iHep cells) to phytochemicals in comparison with hepatoblasoma-derived HepG2 cells and long-term human hepatocytes (LTHHs). The iHep cells expressed typical hepatocyte markers cytochrome P450 3A4 (CYP3A4), hepatocyte nuclear factor 4α, and albumin despite the expression of immature markers α-fetoprotein and cytokeratin 19...
June 11, 2018: Toxicology in Vitro: An International Journal Published in Association with BIBRA
https://www.readbyqxmd.com/read/29902227/modeling-human-early-otic-sensory-cell-development-with-induced-pluripotent-stem-cells
#9
Hanae Lahlou, Alejandra Lopez-Juarez, Arnaud Fontbonne, Emmanuel Nivet, Azel Zine
The inner ear represents a promising system to develop cell-based therapies from human induced pluripotent stem cells (hiPSCs). In the developing ear, Notch signaling plays multiple roles in otic region specification and for cell fate determination. Optimizing hiPSC induction for the generation of appropriate numbers of otic progenitors and derivatives, such as hair cells, may provide an unlimited supply of cells for research and cell-based therapy. In this study, we used monolayer cultures, otic-inducing agents, Notch modulation, and marker expression to track early and otic sensory lineages during hiPSC differentiation...
2018: PloS One
https://www.readbyqxmd.com/read/29901119/crispr-therapeutic-tools-for-complex-genetic-disorders-and-cancer-review
#10
Stella Baliou, Maria Adamaki, Anthony M Kyriakopoulos, Demetrios A Spandidos, Michalis Panagiotidis, Ioannis Christodoulou, Vassilis Zoumpourlis
One of the fundamental discoveries in the field of biology is the ability to modulate the genome and to monitor the functional outputs derived from genomic alterations. In order to unravel new therapeutic options, scientists had initially focused on inducing genetic alterations in primary cells, in established cancer cell lines and mouse models using either RNA interference or cDNA overexpression or various programmable nucleases [zinc finger nucleases (ZNF), transcription activator-like effector nucleases (TALEN)]...
June 6, 2018: International Journal of Oncology
https://www.readbyqxmd.com/read/29899557/a-single-cell-high-content-assay-detects-mitochondrial-dysfunction-in-ipsc-derived-neurons-with-mutations-in-snca
#11
Daniel Little, Christin Luft, Olukunbi Mosaku, Maëlle Lorvellec, Zhi Yao, Sébastien Paillusson, Janos Kriston-Vizi, Sonia Gandhi, Andrey Y Abramov, Robin Ketteler, Michael J Devine, Paul Gissen
Mitochondrial dysfunction is implicated in many neurodegenerative diseases including Parkinson's disease (PD). Induced pluripotent stem cells (iPSCs) provide a unique cell model for studying neurological diseases. We have established a high-content assay that can simultaneously measure mitochondrial function, morphology and cell viability in iPSC-derived dopaminergic neurons. iPSCs from PD patients with mutations in SNCA and unaffected controls were differentiated into dopaminergic neurons, seeded in 384-well plates and stained with the mitochondrial membrane potential dependent dye TMRM, alongside Hoechst-33342 and Calcein-AM...
June 13, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29898922/epigenetic-alterations-mediate-ipsc-normalization-of-dna-repair-expression-and-tnr-stability-in-huntington-s-disease
#12
Peter A Mollica, Martina Zamponi, John A Reid, Deepak K Sharma, Alyson E White, Roy C Ogle, Robert D Bruno, Patrick C Sachs
Huntington's disease (HD) is a rare autosomal dominant neurodegenerative disorder caused by a cytosine-adenine-guanine (CAG) trinucleotide repeat (TNR) expansion within the HTT gene. The mechanisms underlying HD-associated cellular dysfunction during pluripotency and neurodevelopment, are poorly understood. Here we tested the hypothesis that hypomethylation during cellular reprogramming leads to up-regulation of DNA repair genes and stabilization of TNRs in HD cells. We sought to determine how the HD TNR region is affected by global epigenetic changes through cellular reprogramming and early neurodifferentiation...
June 13, 2018: Journal of Cell Science
https://www.readbyqxmd.com/read/29897633/genetics-of-alcohol-use-disorder-a-role-for-induced-pluripotent-stem-cells
#13
Iya Prytkova, Alison Goate, Ronald P Hart, Paul A Slesinger
Alcohol use disorder (AUD) affects millions of people and costs nearly 250 billion dollars annually. Few effective FDA-approved treatments exist, and more are needed. AUDs have a strong heritability, but only a few genes have been identified with a large effect size on disease phenotype. Genome wide association studies (GWASs) have identified common variants with low effect sizes, most of which are in non-coding regions of the genome. Animal models frequently fail to recapitulate key molecular features of neuropsychiatric disease due to the polygenic nature of the disease, partial conservation of coding regions, and significant disparity in non-coding regions...
June 13, 2018: Alcoholism, Clinical and Experimental Research
https://www.readbyqxmd.com/read/29897225/establishment-of-a-human-ipsc-and-nanofiber-based-microphysiological-blood-brain-barrier-system
#14
Dianjun Qi, Shaohua Wu, Haishuang Lin, Mitchell A Kuss, Yuguo Lei, Alexey Krasnoslobodtsev, Shaheen Ahmed, Chi Zhang, Hyung Joon Kim, Peng Jiang, Bin Duan
The blood-brain barrier (BBB) is an active and complex diffusion barrier that separates the circulating blood from the brain and extracellular fluid, regulates nutrient transportation, and provides protection against various toxic compounds and pathogens. Creating an in vitro microphysiological BBB system, particularly with relevant human cell types, will significantly facilitate the research of neuro-pharmaceutical drug delivery, screening, and transport, as well as improve our understanding of pathologies that are due to BBB damage...
June 13, 2018: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/29896658/generation-of-transplantable-retinal-pigmented-epithelial-rpe-cells-for-treatment-of-age-related-macular-degeneration-amd
#15
Harshini Surendran, Reena J Rathod, Rajarshi Pal
Age-related macular degeneration (AMD) is the foremost cause of blindness in people over the age of 60 worldwide. Clinically, this disease starts with distortion in central vision eventually leading to legal blindness. Vision loss has a significant impact on quality of life and incurs a substantial cost to the economy. Furthermore, AMD is a complex and progressive neurodegenerative disorder that triggers visual impairment due to the loss of retinal pigmented epithelium (RPE) and the light-sensitive photoreceptors that they support, protect and provide nutrition...
June 13, 2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29895861/%C3%AE-synuclein-oligomers-interact-with-atp-synthase-and-open-the-permeability-transition-pore-in-parkinson-s-disease
#16
Marthe H R Ludtmann, Plamena R Angelova, Mathew H Horrocks, Minee L Choi, Margarida Rodrigues, Artyom Y Baev, Alexey V Berezhnov, Zhi Yao, Daniel Little, Blerida Banushi, Afnan Saleh Al-Menhali, Rohan T Ranasinghe, Daniel R Whiten, Ratsuda Yapom, Karamjit Singh Dolt, Michael J Devine, Paul Gissen, Tilo Kunath, Morana Jaganjac, Evgeny V Pavlov, David Klenerman, Andrey Y Abramov, Sonia Gandhi
Protein aggregation causes α-synuclein to switch from its physiological role to a pathological toxic gain of function. Under physiological conditions, monomeric α-synuclein improves ATP synthase efficiency. Here, we report that aggregation of monomers generates beta sheet-rich oligomers that localise to the mitochondria in close proximity to several mitochondrial proteins including ATP synthase. Oligomeric α-synuclein impairs complex I-dependent respiration. Oligomers induce selective oxidation of the ATP synthase beta subunit and mitochondrial lipid peroxidation...
June 12, 2018: Nature Communications
https://www.readbyqxmd.com/read/29893509/characterization-of-induced-pluripotent-stem-cell-derived-megakaryocyte-lysates-for-potential-regenerative-applications
#17
Anja Baigger, Dorothee Eicke, Yuliia Yuzefovych, Denys Pogozhykh, Rainer Blasczyk, Constanca Figueiredo
Recently, platelet-derived growth factors present in lysates became an extreme interest in the field of regenerative medicine such as in wound healing and as substitutes to foetal bovine serum in xeno-free cell culture systems. However, the generation of such platelet lysates completely depends on the availability of platelet donors. In this study, the possibility to use in vitro-generated megakaryocytes derived from induced pluripotent stem cells (iPSCs) as a cell source for typical platelet growth factors was investigated...
June 12, 2018: Journal of Cellular and Molecular Medicine
https://www.readbyqxmd.com/read/29893190/metformin-preconditioning-of-human-induced-pluripotent-stem-cell-derived-neural-stem-cells-promotes-their-engraftment-and-improves-post-stroke-regeneration-and-recovery
#18
Fares Ould-Brahim, Sailendra Nath Sarma, Charvi Syal, Kevin Jiaqi Lu, Matthew Seegobin, Anthony Carter, Matthew S Jeffers, Carole Doré, William Stanford, Dale Corbett, Jing Wang
While transplantation of hiPSC-derived neural stem cells (hiPSC-NSCs) shows therapeutic potential in animal stroke models, major concerns for translating hiPSC therapy to the clinic are efficacy and safety. Therefore, there is a demand to develop an optimal strategy to enhance the engraftment and regenerative capacity of transplanted hiPSC-NSCs in order to produce fully differentiated neural cells to replace lost brain tissues. Metformin, an FDA approved drug, is an optimal neuroregenerative agent that not only promotes NSC proliferation but also drives NSC towards differentiation...
June 12, 2018: Stem Cells and Development
https://www.readbyqxmd.com/read/29892062/p53-inhibits-crispr-cas9-engineering-in-human-pluripotent-stem-cells
#19
Robert J Ihry, Kathleen A Worringer, Max R Salick, Elizabeth Frias, Daniel Ho, Kraig Theriault, Sravya Kommineni, Julie Chen, Marie Sondey, Chaoyang Ye, Ranjit Randhawa, Tripti Kulkarni, Zinger Yang, Gregory McAllister, Carsten Russ, John Reece-Hoyes, William Forrester, Gregory R Hoffman, Ricardo Dolmetsch, Ajamete Kaykas
CRISPR/Cas9 has revolutionized our ability to engineer genomes and conduct genome-wide screens in human cells1-3 . Whereas some cell types are amenable to genome engineering, genomes of human pluripotent stem cells (hPSCs) have been difficult to engineer, with reduced efficiencies relative to tumour cell lines or mouse embryonic stem cells3-13 . Here, using hPSC lines with stable integration of Cas9 or transient delivery of Cas9-ribonucleoproteins (RNPs), we achieved an average insertion or deletion (indel) efficiency greater than 80%...
June 11, 2018: Nature Medicine
https://www.readbyqxmd.com/read/29890847/induced-pluripotent-stem-cells-a-powerful-neurodegenerative-disease-modeling-tool-for-mechanism-study-and-drug-discovery
#20
Chia-Yu Chang, Hsiao-Chien Ting, Ching-Ann Liu, Hong-Lin Su, Tzyy-Wen Chiou, Horng-Jyh Harn, Shinn-Zong Lin
Many neurodegenerative diseases are progressive, complex diseases without clear mechanisms or effective treatments. To study the mechanisms underlying these diseases and to develop treatment strategies, a reliable in vitro modeling system is critical. Induced pluripotent stem cells (iPSCs) have the ability to self-renew and possess the differentiation potential to become any kind of adult cell; thus, they may serve as a powerful material for disease modeling. Indeed, patient cell-derived iPSCs can differentiate into specific cell lineages that display the appropriate disease phenotypes and vulnerabilities...
January 1, 2018: Cell Transplantation
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