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https://www.readbyqxmd.com/read/28435121/proteome-analysis-of-human-embryonic-stem-cells-organelles
#1
Faezeh Shekari, Hossein Nezari, Mehran Rezaei Larijani, Chia-Li Han, Hossein Baharvand, Yu-Ju Chen, Ghasem Hosseini Salekdeh
As the functions of proteins are associated with their cellular localization, the comprehensive sub-cellular proteome knowledge of human embryonic stem cells (hESCs) is indispensable for ensuring a therapeutic effect. Here, we have utilized a sub-cellular proteomics approach to analyze the localization of proteins in the nucleus, mitochondria, crude membrane, cytoplasm, heavy and light microsomes. Out of 2002 reproducibly identified proteins, we detected 762 proteins in a single organelle whereas 160 proteins were found in all sub-cellular fractions...
April 20, 2017: Journal of Proteomics
https://www.readbyqxmd.com/read/28433654/high-molecular-weight-fgf2-isoforms-demonstrate-canonical-receptor-mediated-activity-and-support-human-embryonic-stem-cell-self-renewal
#2
Denis Kole, Alexandra Grella, David Dolivo, Lucia Shumaker, William Hermans, Tanja Dominko
Basic fibroblast growth factor (FGF2) is a highly pleiotropic member of a large family of growth factors with a broad range of activities, including mitogenesis and angiogenesis (Ornitz et al., 1996; Zhang et al., 2006), and it is known to be essential for maintenance of balance between survival, proliferation, and self-renewal in human pluripotent stem cells (Eiselleova et al., 2009; Zoumaro-Djayoon et al., 2011). A single FGF2 transcript can be translated into five FGF2 protein isoforms, an 18kDa low molecular weight (LMW) isoform and four larger high molecular weight (HMW) isoforms (Arese et al...
April 18, 2017: Stem Cell Research
https://www.readbyqxmd.com/read/28433627/docosahexaenoic-acid-promotes-differentiation-of-photoreceptor-cells-in-three-dimensional-neural-retinas
#3
Eisuke Arai, Vipul M Parmar, Bhubanananda Sahu, Lindsay Perusek, Tanu Parmar, Akiko Maeda
Retinal tissues generated from human pluripotent stem cells can be an excellent tool for investigating pathogenesis of retinal diseases and developing new pharmacologic therapies. Moreover, patient derived retinal tissues could allow for retinal transplantation therapy for degenerative retinal diseases. However, obtaining retinal tissues with matured photoreceptor outer segments, which are essential for photoreceptor functions, is currently challenging. Here we investigated the effects of docosahexaenoic acid (DHA) for maturation of photoreceptor outer segments at the late stage and visual chromophore analog, 9-cis-retinal for the early stage of differentiation to three-dimensional (3D)-retinal tissues from human embryonic stem cells (hESCs), respectively...
April 19, 2017: Neuroscience Research
https://www.readbyqxmd.com/read/28433626/thrombin-impairs-human-endometrial-endothelial-angiogenesis-implications-for-progestin-only-contraceptive-induced-abnormal-uterine-bleeding
#4
John P Shapiro, Ozlem Guzeloglu-Kayisli, Umit A Kayisli, Nihan Semerci, S Joseph Huang, Sefa Arlier, Kellie Larsen, Paolo Fadda, Frederick Schatz, Charles J Lockwood
OBJECTIVE: Progestin-only contraceptives induce abnormal uterine bleeding, accompanied by pro-thrombin leakage from dilated endometrial microvessels and increased thrombin generation by human endometrial stromal cell (HESC)-expressed tissue factor. Initial studies of the thrombin-treated HESC secretome identified elevated levels of cleaved chondroitin sulfate proteoglycan 4 (CSPG4), impairing pericyte-endothelial interactions. Thus, we investigated direct and CSPG4-mediated effects of thrombin in eliciting abnormal uterine bleeding by disrupting endometrial angiogenesis...
April 19, 2017: Contraception
https://www.readbyqxmd.com/read/28430877/the-accessible-chromatin-landscape-during-conversion-of-human-embryonic-stem-cells-to-trophoblast-by-bmp4
#5
Yajun Liu, Dewu Ding, Hongde Liu, Xiao Sun
Human embryonic stem cells (hESC) exposed to the growth factor bone morphogenic protein 4 (BMP4) in the absence of FGF2 have been used as a model to study the development of placental development. However, little is known about the cis-regulatory mechanisms underlying this important process. In this study, we used the public available chromatin accessibility data of hESC H1 cells and BMP4-induced trophoblast (TB)cell lines to identify DNase I hypersensitive sites (DHSs) in the two cell lines, as well as the transcription factor binding sites within the DHSs...
April 18, 2017: Biology of Reproduction
https://www.readbyqxmd.com/read/28429706/direct-comparison-of-distinct-naive-pluripotent-states-in-human-embryonic-stem-cells
#6
S Warrier, M Van der Jeught, G Duggal, L Tilleman, E Sutherland, J Taelman, M Popovic, S Lierman, S Chuva De Sousa Lopes, A Van Soom, L Peelman, F Van Nieuwerburgh, D I M De Coninck, B Menten, P Mestdagh, J Van de Sompele, D Deforce, P De Sutter, B Heindryckx
Until recently, human embryonic stem cells (hESCs) were shown to exist in a state of primed pluripotency, while mouse embryonic stem cells (mESCs) display a naive or primed pluripotent state. Here we show the rapid conversion of in-house-derived primed hESCs on mouse embryonic feeder layer (MEF) to a naive state within 5-6 days in naive conversion media (NCM-MEF), 6-10 days in naive human stem cell media (NHSM-MEF) and 14-20 days using the reverse-toggle protocol (RT-MEF). We further observe enhanced unbiased lineage-specific differentiation potential of naive hESCs converted in NCM-MEF, however, all naive hESCs fail to differentiate towards functional cell types...
April 21, 2017: Nature Communications
https://www.readbyqxmd.com/read/28413007/a-homozygous-keap1-knockout-human-embryonic-stem-cell-line-generated-using-crispr-cas9-mediates-gene-targeting
#7
So-Jung Kim, Omer Habib, Jin-Soo Kim, Hyo-Won Han, Soo Kyung Koo, Jung-Hyun Kim
Kelch-like ECH-associated protein 1 (keap1) is a cysteine-rich protein that interacts with transcription factor Nrf2 in a redox-sensitive manner, leading to the degradation of Nrf2 (Kim et al., 2014a). Disruption of Keap1 results in the induction of Nrf2-related signaling pathways involving the expression of a set of anti-oxidant and anti-inflammatory genes. We generated biallelic mutants of the Keap1 gene using a CRISPR-Cas9 genome editing method in the H9 human embryonic stem cell (hESC). The Keap1 homozygous-knockout H9 cell line retained normal morphology, gene expression, and in vivo differentiation potential...
March 2017: Stem Cell Research
https://www.readbyqxmd.com/read/28413005/generation-of-a-nrf2-homozygous-knockout-human-embryonic-stem-cell-line-using-crispr-cas9
#8
So-Jung Kim, Omer Habib, Jin-Soo Kim, Hyo-Won Han, Soo Kyung Koo, Jung-Hyun Kim
Nuclear factor erythroid 2-related factor 2 (NFE2L2 or Nrf2) is a well-known transcription factor that regulates the expression of a large number of anti-oxidant genes in mammalian cells (J.H. Kim et al., 2014). Here, we generated a homozygous Nrf2 knockout human embryonic stem cell (hESC) line, H9Nrf2KO-A13, using the CRISPR/Cas9 genome editing method. The Nrf2 homozygous knockout H9 cell line maintains pluripotency, differentiation potential into three germ layers, and a normal karyotype.
March 2017: Stem Cell Research
https://www.readbyqxmd.com/read/28412746/kr%C3%A3-ppel-like-factor-4-klf4-regulates-the-mir-183-96-182-cluster-under-physiologic-and-pathologic-conditions
#9
Miguel F Segura, Luz Jubierre, SiDe Li, Aroa Soriano, Lisa Koetz, Avital Gaziel-Sovran, Marc Masanas, Kevin Kleffman, John F Dankert, Martin J Walsh, Eva Hernando
MicroRNAs (miRNAs) are a class of endogenous non-coding small RNAs that post-transcriptionally control the translation and stability of target mRNAs in a sequence-dependent manner. MiRNAs are essential for key cellular processes including proliferation, differentiation, cell death and metabolism, among others. Consequently, alterations of miRNA expression contribute to developmental defects and a myriad of diseases.The expression of miRNAs can be altered by several mechanisms including gene copy number alterations, aberrant DNA methylation, defects of the miRNA processing machinery or unscheduled expression of transcription factors...
February 17, 2017: Oncotarget
https://www.readbyqxmd.com/read/28400986/combining-injectable-plasma-scaffold-with-mesenchymal-stem-stromal-cells-for-repairing-infarct-cavity-after-ischemic-stroke
#10
Hongxia Zhang, Fen Sun, Jixian Wang, Luokun Xie, Chenqi Yang, Mengxiong Pan, Bei Shao, Guo-Yuan Yang, Shao-Hua Yang, Qichuan ZhuGe, Kunlin Jin
Stroke survivors are typically left with structural brain damage and associated functional impairment in the chronic phase of injury, for which few therapeutic options exist. We reported previously that transplantation of human embryonic stem cell (hESC)-derived neural stem cells together with Matrigel scaffolding into the brains of rats after focal ischemia reduced infarct volume and improved neurobehavioral performance. Matrigel is a gelatinous protein mixture extracted from mouse sarcoma cells, thus would not be approved for use as a scaffold clinically...
April 2017: Aging and Disease
https://www.readbyqxmd.com/read/28392220/modeling-developmental-and-tumorigenic-aspects-of-trilateral-retinoblastoma-via-human-embryonic-stem-cells
#11
Yishai Avior, Elyad Lezmi, Dorit Yanuka, Nissim Benvenisty
Human embryonic stem cells (hESCs) provide a platform for studying human development and understanding mechanisms underlying diseases. Retinoblastoma-1 (RB1) is a key regulator of cell cycling, of which biallelic inactivation initiates retinoblastoma, the most common congenital intraocular malignancy. We developed a model to study the role of RB1 in early development and tumor formation by generating RB1-null hESCs using CRISPR/Cas9. RB1(-/-) hESCs initiated extremely large teratomas, with neural expansions similar to those of trilateral retinoblastoma tumors, in which retinoblastoma is accompanied by intracranial neural tumors...
March 22, 2017: Stem Cell Reports
https://www.readbyqxmd.com/read/28387893/regenerative-medicine-in-cardiovascular-diseases-an-update
#12
Y Cao, P-Y Zhang
The unique ability of regenerative medicine to differentiate into any cell of the three germ layers (endoderm, ectoderm, and mesoderm) is of immense clinical importance. They have an unique capacity for unlimited self-renewal. Furthermore, pluripotent stem cells (PSC), including human embryonic (hESC) and induced pluripotent stem cells (hiPSCs), hold great potential as an unlimited source of functional as well as transplantable cells applications. Specifically, in the context of cardiovascular and ischemic diseases, it is believed that hESC-derived endothelial cells (hESC- ECs) could be used to stimulate angiogenesis or vasculogenesis in ischemic tissues, thereby restoring blood supply to the affected area...
March 2017: European Review for Medical and Pharmacological Sciences
https://www.readbyqxmd.com/read/28387239/survival-of-human-embryonic-stem-cells-implanted-in-the-guinea-pig-auditory-epithelium
#13
Min Young Lee, Sandra Hackelberg, Kari L Green, Kelly G Lunghamer, Takaomi Kurioka, Benjamin R Loomis, Donald L Swiderski, R Keith Duncan, Yehoash Raphael
Hair cells in the mature cochlea cannot spontaneously regenerate. One potential approach for restoring hair cells is stem cell therapy. However, when cells are transplanted into scala media (SM) of the cochlea, they promptly die due to the high potassium concentration. We previously described a method for conditioning the SM to make it more hospitable to implanted cells and showed that HeLa cells could survive for up to a week using this method. Here, we evaluated the survival of human embryonic stem cells (hESC) constitutively expressing GFP (H9 Cre-LoxP) in deaf guinea pig cochleae that were pre-conditioned to reduce potassium levels...
April 7, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28382404/prediction-of-hepatotoxicity-for-drugs-using-human-pluripotent-stem-cell-derived-hepatocytes
#14
Jong Hyun Kim, Min Wang, Jaehun Lee, Han-Jin Park, Chungseong Han, Hee Su Hong, Jeong Seong Kim, Geun Ho An, Kijung Park, Hee-Kyung Park, Shi Feng Zhu, Xiao-Bo Sun, Jong-Hoon Kim, Dong-Hun Woo
Drug-induced liver toxicity is a main reason for withdrawals of new drugs in late clinical phases and post-launch of the drugs. Thus, hepatotoxicity screening of drug candidates in pre-clinical stage is important for reducing drug attrition rates during the clinical development process. Here, we show commercially available hepatocytes that could be used for early toxicity evaluation of drug candidates. From our hepatic differentiation technology, we obtained highly pure (≥98%) hepatocytes from human embryonic stem cells (hESCs) having mature phenotypes and similar gene expression profiles with those of primary human tissues...
April 5, 2017: Cell Biology and Toxicology
https://www.readbyqxmd.com/read/28380361/efficient-generation-of-glucose-responsive-beta-cells-from-isolated-gp2-human-pancreatic-progenitors
#15
Jacqueline Ameri, Rehannah Borup, Christy Prawiro, Cyrille Ramond, Karen A Schachter, Raphael Scharfmann, Henrik Semb
Stem cell-based therapy for type 1 diabetes would benefit from implementation of a cell purification step at the pancreatic endoderm stage. This would increase the safety of the final cell product, allow the establishment of an intermediate-stage stem cell bank, and provide a means for upscaling β cell manufacturing. Comparative gene expression analysis revealed glycoprotein 2 (GP2) as a specific cell surface marker for isolating pancreatic endoderm cells (PECs) from differentiated hESCs and human fetal pancreas...
April 4, 2017: Cell Reports
https://www.readbyqxmd.com/read/28377585/messenger-rnas-localized-to-distal-projections-of-human-stem-cell-derived-neurons
#16
Rebecca L Bigler, Joyce W Kamande, Raluca Dumitru, Mark Niedringhaus, Anne Marion Taylor
The identification of mRNAs in distal projections of model organisms has led to the discovery of multiple proteins that are locally synthesized for functional roles such as axon guidance, injury signaling and regeneration. The extent to which local protein synthesis is conserved in human neurons is unknown. Here we used compartmentalized microfluidic chambers to characterize the transcriptome of distal projections of human embryonic stem cells differentiated using a protocol which enriched for glutamatergic neurons (hESC-neurons)...
April 4, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28373677/dynamics-of-single-human-embryonic-stem-cells-and-their-pairs-a-quantitative-analysis
#17
L E Wadkin, L F Elliot, I Neganova, N G Parker, V Chichagova, G Swan, A Laude, M Lako, A Shukurov
Numerous biological approaches are available to characterise the mechanisms which govern the formation of human embryonic stem cell (hESC) colonies. To understand how the kinematics of single and pairs of hESCs impact colony formation, we study their mobility characteristics using time-lapse imaging. We perform a detailed statistical analysis of their speed, survival, directionality, distance travelled and diffusivity. We confirm that single and pairs of cells migrate as a diffusive random walk for at least 7 hours of evolution...
April 3, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28370937/the-n-glycome-of-embryonic-stem-cells-and-thereof-differentiated-hepatic-cells
#18
Houda Montacir, Nora Freyer, Fanny Knoespel, Thomas Urbaniak, Tereza Dedova, Markus Berger, Georg Damm, Rudolf Tauber, Katrin Zeilinger, Veronique Blanchard
Human embryonic stem cells (hESC) are unspecialized cells that are used in regenerative medicine. In addition, the have been proposed as an appropriate alternative source of hepatocytes. In this work, hESC were differentiated into definitive endodermal cells (DEC) then hepatocyte-like cells (HLC). Their N-glycome was measured and compared also with the one of primary hepatocytes. hESC contained a high amount of high-mannose N-glycans. In contrast, complex-type N-glycans such as asialylated or monosialylated biantennary and triantennary N-glycans were dominant in HLC and fully-galactosylated structures were much more abundant than in undifferentiated hESC...
March 31, 2017: Chembiochem: a European Journal of Chemical Biology
https://www.readbyqxmd.com/read/28362406/efficient-differentiation-of-pluripotent-stem-cells-to-nkx6-1-pancreatic-progenitors
#19
Emily C McGaugh, M Cristina Nostro
Pluripotent stem cells have the ability to self renew and differentiate to multiple lineages, making them an attractive source for the generation of pancreatic progenitor cells that can be used for the study of and future treatment of diabetes. This article outlines a four-stage differentiation protocol designed to generate pancreatic progenitor cells from human embryonic stem cells (hESCs). This protocol can be applied to a number of human pluripotent stem cell (hPSC) lines. The approach taken to generate pancreatic progenitor cells is to differentiate hESCs to accurately model key stages of pancreatic development...
March 7, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28359310/increased-kr%C3%A3-ppel-like-factor-12-in-recurrent-implantation-failure-impairs-endometrial-decidualization-by-repressing-nur77-expression
#20
Chenyang Huang, Yue Jiang, Jianjun Zhou, Qiang Yan, Ruiwei Jiang, Xi Cheng, Jun Xing, Lijun Ding, Jianxin Sun, Guijun Yan, Haixiang Sun
BACKGROUND: Decidualization is a prerequisite for successful implantation and the establishment of pregnancy. A critical role of impaired decidualization in subfertility has been established. In human endometrial stromal cells (hESCs), Krüppel-like factor 12 (KLF12) and Nur77 are novel regulators of decidualization. We investigated whether KLF12 impaired the decidualization of hESCs in recurrent implantation failure (RIF) patients. METHODS: Endometrial tissues and hESCs were collected from RIF patients (n = 34) and fertile controls (n = 30) for in vitro analysis...
March 31, 2017: Reproductive Biology and Endocrinology: RB&E
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