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https://www.readbyqxmd.com/read/27911036/properties-of-embryoid-bodies
#1
REVIEW
Joshua M Brickman, Palle Serup
Embryoid bodies (EBs) have been popular in vitro differentiation models for pluripotent stem cells for more than five decades. Initially, defined as aggregates formed by embryonal carcinoma cells, EBs gained more prominence after the derivation of karyotypically normal embryonic stem cells from early mouse blastocysts. In many cases, formation of EBs constitutes an important initial step in directed differentiation protocols aimed at generated specific cell types from undifferentiated stem cells. Indeed state-of-the-art protocols for directed differentiation of cardiomyocytes still rely on this initial EB step...
December 2, 2016: Wiley Interdisciplinary Reviews. Developmental Biology
https://www.readbyqxmd.com/read/27904699/generation-of-induced-pluripotent-stem-cells-with-high-efficiency-from-human-embryonic-renal-cortical-cells
#2
Ling Yao, Ruifang Chen, Pu Wang, Qi Zhang, Hailiang Tang, Huaping Sun
Reprogramming of somatic cells into induced pluripotent stem cells (iPSCs) emerges as a prospective therapeutic angle in regenerative medicine and a tool for drug screening. Although increasing numbers of iPSCs from different sources have been generated, there has been limited progress in yield of iPSC. Here, we show that four Yamanaka factors Oct4, Sox2, Klf4 and c-Myc can convert human embryonic renal cortical cells (hERCCs) to pluripotent stem cells with a roughly 40-fold higher reprogramming efficiency compared with that of adult human dermal fibroblasts...
2016: American Journal of Translational Research
https://www.readbyqxmd.com/read/27903111/electric-stimulation-enhances-cardiac-differentiation-of-human-induced-pluripotent-stem-cells-for-myocardial-infarction-therapy
#3
Ruilian Ma, Jialiang Liang, Wei Huang, Linlin Guo, WenFeng Cai, Lei Wang, Christian Paul, Huang-Tian Yang, Ha Won Kim, Yigang Wang
AIM: Electrical stimulation (EleS) can promote cardiac differentiation, but the underlying mechanism is not well known. This study investigated the effect of EleS on cardiomyocyte (CM) differentiation of human induced pluripotent stem cells (hiPSCs) and evaluated the therapeutic effects for the treatment of myocardial infarction (MI). RESULTS: Cardiac differentiation of hiPSCs was induced with EleS after embryoid body formation. Spontaneously beating hiPSCs were observed as early at 2 days when treated with EleS as compared to control treatment...
November 30, 2016: Antioxidants & Redox Signaling
https://www.readbyqxmd.com/read/27883036/the-mevalonate-pathway-regulates-primitive-streak-formation-via-protein-farnesylation
#4
Yoshimi Okamoto-Uchida, Ruoxing Yu, Norio Miyamura, Norie Arima, Mari Ishigami-Yuasa, Hiroyuki Kagechika, Suguru Yoshida, Takamitsu Hosoya, Makiko Nawa, Takeshi Kasama, Yoichi Asaoka, Reiner Wimmer Alois, Ulrich Elling, Josef M Penninger, Sachiko Nishina, Noriyuki Azuma, Hiroshi Nishina
The primitive streak in peri-implantation embryos forms the mesoderm and endoderm and controls cell differentiation. The metabolic cues regulating primitive streak formation remain largely unknown. Here we utilised a mouse embryonic stem (ES) cell differentiation system and a library of well-characterised drugs to identify these metabolic factors. We found that statins, which inhibit the mevalonate metabolic pathway, suppressed primitive streak formation in vitro and in vivo. Using metabolomics and pharmacologic approaches we identified the downstream signalling pathway of mevalonate and revealed that primitive streak formation requires protein farnesylation but not cholesterol synthesis...
November 24, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27873029/induction-of-pluripotency-in-long-term-cryopreserved-human-neonatal-fibroblasts-in-feeder-free-condition
#5
Maria Csobonyeiova, Lubica Krajciova, Andreas Nicodemou, Stefan Polak, Lubos Danisovic
A novel approach for stem cell generation is the attempt to induce conversion of the adult somatic cells into pluripotent stem cells so called induced pluripotent stem cells (iPSCs) by introducing specific transcription factors. iPSCs have two essential cell characteristics, they are pluripotent and posses long term cell-renewal capacity. Additionally, iPSCs can be derived from patient-specific somatic cells, thus bypassing ethical and immunological issues. The aim of our study was to reprogram long-term cryopreserved human neonatal fibroblasts by new method using lipid nano-particle technology (Lipofectamine 3000 reagent transfection system) in combination with Epi 5 reprogramming vectors...
November 21, 2016: Cell and Tissue Banking
https://www.readbyqxmd.com/read/27870307/abrogation-of-gap-junctional-communication-in-es-cells-results-in-a-disruption-of-primitive-endoderm-formation-in-embryoid-bodies
#6
Philipp Wörsdörfer, Felicitas Bosen, Martina Gebhardt, Nicole Russ, Katrin Zimmermann, David Komla Kessie, Thileepan Sekaran, Angela Egert, Süleyman Ergün, Hubert Schorle, Alexander Pfeifer, Frank Edenhofer, Klaus Willecke
Gap junctional intercellular communication (GJIC) has been suggested to be involved in early embryonic development but the actual functional role remained elusive. Connexin (Cx) 43 and Cx45 are co-expressed in embryonic stem (ES) cells, form gap junctions and are considered to exhibit adhesive function and/or to contribute to the establishment of defined communication compartments. Here we describe the generation of Cx43/Cx45-double deficient mouse ES cells to achieve almost complete breakdown of GJIC. Cre-loxP induced deletion of both, Cx43 and Cx45, results in a block of differentiation in embryoid bodies without affecting pluripotency marker expression and proliferation in ES cells...
November 21, 2016: Stem Cells
https://www.readbyqxmd.com/read/27864696/generation-and-differentiation-of-induced-pluripotent-stem-cells-reveal-ankylosing-spondylitis-risk-gene-expression-in-bone-progenitors
#7
Gerlinde Layh-Schmitt, Shajia Lu, Fatemeh Navid, Stephen R Brooks, Emily Lazowick, Kathryn M Davis, Cristina Montagna, Massimo Gadina, Robert A Colbert
Axial spondyloarthritis (axSpA), which encompasses ankylosing spondylitis, is a complex genetic disease. Aberrant bone formation is a key feature of pathogenesis that can lead to ankylosis of the spine. Our objective is to determine, whether genes whose variants confer susceptibility to AS are expressed in bone progenitors like mesenchymal stem cells (MSCs). Since MSCs from bone marrow is difficult to obtain, we first examined, whether MSCs can be derived from induced pluripotent stem cells (iPSCs). Dermal fibroblasts of two axSpA patients and one healthy control were reprogrammed into iPSCs using a Sendai virus vector encoding pluripotency genes...
November 18, 2016: Clinical Rheumatology
https://www.readbyqxmd.com/read/27864336/bmp4-promotes-mouse-ips-cell-differentiation-to-male-germ-cells-via-smad1-5-gata4-id1-and-id2
#8
Shi Yang, Qingqing Yuan, Minghui Niu, Jingmei Hou, Zijue Zhu, Min Sun, Zheng Li, Zuping He
Generation of male germ cells from pluripotent cells could provide male gametes for treating male infertility and offer an ideal model for unveiling molecular mechanisms of spermatogenesis. However, the influence and exact molecular mechanisms, especially downstream effectors of BMP4 signaling pathways, in male germ cell differentiation of the induce pluripotent stem (iPS) cells, remain unknown. This study was designed to explore the role and mechanism of BMP4 signaling in the differentiation of mouse induced pluripotent stem (iPS) cells to male germ cells...
November 18, 2016: Reproduction: the Official Journal of the Society for the Study of Fertility
https://www.readbyqxmd.com/read/27846905/rapamycin-regulates-autophagy-and-cell-adhesion-in-induced-pluripotent-stem-cells
#9
Areechun Sotthibundhu, Katya McDonagh, Alexander von Kriegsheim, Amaya Garcia-Munoz, Agnieszka Klawiter, Kerry Thompson, Kapil Dev Chauhan, Janusz Krawczyk, Veronica McInerney, Peter Dockery, Michael J Devine, Tilo Kunath, Frank Barry, Timothy O'Brien, Sanbing Shen
BACKGROUND: Cellular reprogramming is a stressful process, which requires cells to engulf somatic features and produce and maintain stemness machineries. Autophagy is a process to degrade unwanted proteins and is required for the derivation of induced pluripotent stem cells (iPSCs). However, the role of autophagy during iPSC maintenance remains undefined. METHODS: Human iPSCs were investigated by microscopy, immunofluorescence, and immunoblotting to detect autophagy machinery...
November 15, 2016: Stem Cell Research & Therapy
https://www.readbyqxmd.com/read/27845273/real-time-and-non-invasive-monitoring-of-embryonic-stem-cell-survival-during-the-development-of-embryoid-bodies-with-smart-nanosensor
#10
Jiayin Fu, Christian Wiraja, Ruiqi Chong, Chenjie Xu, Dong-An Wang
: Embryonic stem cells (ESCs)-derived embryoid body (EB) is a powerful model for the study of early embryonic development and the discovery of therapeutics for tissue regeneration. This article reports a smart nanosensor platform for labeling and tracking the survival and distribution of ESCs during the EB development in a real-time and non-invasive way. Compared with the cell tracker (i.e. DiO) and the green fluorescent protein (GFP), nanosensors provide the homogenous and highly-efficient ESC labeling...
November 11, 2016: Acta Biomaterialia
https://www.readbyqxmd.com/read/27834666/proliferation-in-culture-of-primordial-germ-cells-derived-from-embryonic-stem-cell-induction-by-retinoic-acid
#11
Zohreh Makoolati, Mansoureh Movahedin, Mehdi Forouzandeh-Moghadam
An in vitro system that supports primordial germ cells (PGCs) survival and proliferation is useful for enhancement of these cells and efficient transplantation in infertility disorders. One approach is cultivation of PGCs under proper conditions that allow self-renewal and proliferation of PGCs. For this purpose, we compared the effects of different concentrations of retinoic acid (RA), and the effect of PGCs co-culture with STO cells on the proliferation of embryonic stem cells (ESCs)- derived PGCs. One-day-old embryoid body (EB) was cultured for 4 days in simple culture system at the presence of 5 ng/ml BMP4 (SCB group) for PGC induction...
November 10, 2016: Bioscience Reports
https://www.readbyqxmd.com/read/27826746/protein-tyrosine-phosphatase-1b-ptp1b-is-required-for-cardiac-lineage-differentiation-of-mouse-embryonic-stem-cells
#12
Zahra Shokati Eshkiki, Mohammad Hossein Ghahremani, Parisa Shabani, Sattar Gorgani Firuzjaee, Asie Sadeghi, Hossein Ghanbarian, Reza Meshkani
Protein tyrosine phosphatase 1B (PTP1B) has been shown to regulate multiple cellular events such as differentiation, cell growth, and proliferation; however, the role of PTP1B in differentiation of embryonic stem (ES) cells into cardiomyocytes remains unexplored. In the present study, we investigated the effects of PTP1B inhibition on differentiation of ES cells into cardiomyocytes. PTP1B mRNA and protein levels were increased during the differentiation of ES cells into cardiomyocytes. Accordingly, a stable ES cell line expressing PTP1B shRNA was established...
November 8, 2016: Molecular and Cellular Biochemistry
https://www.readbyqxmd.com/read/27826002/3d-culture-of-human-pluripotent-stem-cells-in-rgd-alginate-hydrogel-improves-retinal-tissue-development
#13
Nicola C Hunt, Dean Hallam, Ayesha Karimi, Carla B Mellough, Jinju Chen, David H W Steel, Majlinda Lako
: No treatments exist to effectively treat many retinal diseases. Retinal pigmented epithelium (RPE) and neural retina can be generated from human embryonic stem cells/induced pluripotent stem cells (hESCs/hiPSCs). The efficacy of current protocols is, however, limited. It was hypothesised that generation of laminated neural retina and/or RPE from hiPSCs/hESCs could be enhanced by 3D culture in hydrogels. hiPSC- and hESC-derived embryoid bodies (EBs) were encapsulated in 0.5% RGD-alginate; 1% RGD-alginate; hyaluronic acid (HA) or HA/gelatin hydrogels and maintained until day 45...
November 5, 2016: Acta Biomaterialia
https://www.readbyqxmd.com/read/27812821/early-retinal-differentiation-of-human-pluripotent-stem-cells-in-microwell-suspension-cultures
#14
Vishal S Sharma, Rana Khalife, Rui Tostoes, Leonard Leung, Rose Kinsella, Ludmilla Ruban, Farlan S Veraitch
OBJECTIVE: To develop a microwell suspension platform for the adaption of attached stem cell differentiation protocols into mixed suspension culture. RESULTS: We adapted an adherent protocol for the retinal differentiation of human induced pluripotent stem cells (hiPSCs) using a two-step protocol. Establishing the optimum embryoid body (EB) starting size and shaking speed resulted in the translation of the original adherent process into suspension culture. Embryoid bodies expanded in size as the culture progressed resulting in the expression of characteristic markers of early (Rx, Six and Otx2) and late (Crx, Nrl and Rhodopsin) retinal differentiation...
November 3, 2016: Biotechnology Letters
https://www.readbyqxmd.com/read/27802323/derivation-of-patient-specific-pluripotent-stem-cells-using-clinically-discarded-cumulus-cells
#15
Wei-Fang Chang, Yuh-Ming Hwu, Jie Xu, Chen-Ju Lin, Sheng-Wen Wang, An-Sheng Cheng, Jean Lu, Chung-Hao Lu, Li-Ying Sung
Induced pluripotent stem cells (iPSCs) are powerful tools for basic and translational research, as well as regenerative medicine. In routine human in vitro fertilization (IVF) practices, cumulus cells (CCs) are discarded, representing a potential source of biological materials for regenerative medicine. In this study, we derived patient-specific iPSCs using CCs from human infertility clinics for the first time. The human cumulus cell derived iPSCs (hc-iPSCs) were characterized for growth, karyotype, expression of pluripotency genes, and were subjected to embryoid bodies (EBs) and teratoma assays to evaluate their differentiation capacity...
2016: PloS One
https://www.readbyqxmd.com/read/27790820/role-of-alpha-and-beta-adrenergic-receptors-in-cardiomyocyte-differentiation-from-murine-induced-pluripotent-stem-cells
#16
Xiao-Li Li, Di Zeng, Yan Chen, Lu Ding, Wen-Ju Li, Ting Wei, Dong-Bo Ou, Song Yan, Bin Wang, Qiang-Sun Zheng
OBJECTIVES: Induced pluripotent stem cell (iPSC)-derived cardiomyocytes are a promising source of cells for regenerative heart disease therapies, but progress towards their use has been limited by their low differentiation efficiency and high cellular heterogeneity. Previous studies have demonstrated expression of adrenergic receptors (ARs) in stem cells after differentiation; however, roles of ARs in fate specification of stem cells, particularly in cardiomyocyte differentiation and development, have not been characterized...
October 27, 2016: Cell Proliferation
https://www.readbyqxmd.com/read/27789412/characterization-of-dermal-fibroblast-derived-ipscs-from-a-patient-with-high-grade-steatosis
#17
Marie-Ann Kawala, Martina Bohndorf, Nina Graffmann, Wasco Wruck, Kurt Zatloukal, James Adjaye
Primary fibroblasts from a high grade steatosis patient were reprogrammed by transduction of retroviruses OCT4, SOX2, c-MYC and KLF4. IPSCs were characterized by immunocytochemistry, embryoid body-formation, DNA-fingerprint, karyotype analysis and comparative transcriptome analyses with the human embryonic stem cell line H1 revealed a Pearsons correlation coefficient of 0.9287. Resource table.
October 19, 2016: Stem Cell Research
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
#18
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/27770834/preclinical-study-of-mouse-pluripotent-parthenogenetic-embryonic-stem-cell-derivatives-for-the-construction-of-tissue-engineered-skin-equivalent
#19
Yang Rao, Jihong Cui, Lu Yin, Wei Liu, Wenguang Liu, Mei Sun, Xingrong Yan, Ling Wang, Fulin Chen
BACKGROUND: Embryonic stem cell (ESC) derivatives hold great promise for the construction of tissue-engineered skin equivalents (TESE). However, harvesting of ESCs destroys viable embryos and may lead to political and ethical concerns over their application. In the current study, we directed mouse parthenogenetic embryonic stem cells (pESCs) to differentiate into fibroblasts, constructed TESE, and evaluated its function in vivo. METHODS: The stemness marker expression and the pluripotent differentiation ability of pESCs were tested...
October 22, 2016: Stem Cell Research & Therapy
https://www.readbyqxmd.com/read/27748754/differentiation-of-human-embryonic-stem-cells-to-hoxa-hemogenic-vasculature-that-resembles-the-aorta-gonad-mesonephros
#20
Elizabeth S Ng, Lisa Azzola, Freya F Bruveris, Vincenzo Calvanese, Belinda Phipson, Katerina Vlahos, Claire Hirst, Vanta J Jokubaitis, Qing C Yu, Jovana Maksimovic, Simone Liebscher, Vania Januar, Zhen Zhang, Brenda Williams, Aude Conscience, Jennifer Durnall, Steven Jackson, Magdaline Costa, David Elliott, David N Haylock, Susan K Nilsson, Richard Saffery, Katja Schenke-Layland, Alicia Oshlack, Hanna K A Mikkola, Edouard G Stanley, Andrew G Elefanty
The ability to generate hematopoietic stem cells from human pluripotent cells would enable many biomedical applications. We find that hematopoietic CD34(+) cells in spin embryoid bodies derived from human embryonic stem cells (hESCs) lack HOXA expression compared with repopulation-competent human cord blood CD34(+) cells, indicating incorrect mesoderm patterning. Using reporter hESC lines to track the endothelial (SOX17) to hematopoietic (RUNX1C) transition that occurs in development, we show that simultaneous modulation of WNT and ACTIVIN signaling yields CD34(+) hematopoietic cells with HOXA expression that more closely resembles that of cord blood...
October 17, 2016: Nature Biotechnology
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