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https://www.readbyqxmd.com/read/28213969/highly-efficient-neural-conversion-of-human-pluripotent-stem-cells-in-adherent-and-animal-free-conditions
#1
Dunja Lukovic, Andrea Diez Lloret, Petra Stojkovic, Daniel Rodríguez-Martínez, Maria Amparo Perez Arago, Francisco Javier Rodriguez-Jimenez, Patricia González-Rodríguez, José López-Barneo, Eva Sykova, Pavla Jendelova, Jelena Kostic, Victoria Moreno-Manzano, Miodrag Stojkovic, Shomi S Bhattacharya, Slaven Erceg
Neural differentiation of human embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs) can produce a valuable and robust source of human neural cell subtypes, holding great promise for the study of neurogenesis and development, and for treating neurological diseases. However, current hESCs and hiPSCs neural differentiation protocols require either animal factors or embryoid body formation, which decreases efficiency and yield, and strongly limits medical applications. Here we develop a simple, animal-free protocol for neural conversion of both hESCs and hiPSCs in adherent culture conditions...
February 18, 2017: Stem Cells Translational Medicine
https://www.readbyqxmd.com/read/28193206/inducing-goat-pluripotent-stem-cells-with-four-transcription-factor-mrnas-that-activate-endogenous-promoters
#2
Hao Chen, Qisheng Zuo, Yingjie Wang, Jiuzhou Song, Huilin Yang, Yani Zhang, Bichun Li
BACKGROUND: Traditional approaches for generating goat pluripotent stem cells (iPSCs) suffer from complexity and low preparation efficiency. Therefore, we tried to derive goat iPSCs with a new method by transfecting exogenous Oct4, Sox2, Klf4 and c-Myc mRNAs into goat embryonic fibroblasts (GEFs), and explore the mechanisms regarding the transcription regulation of the reprogramming factors in goat iPSCs induction. RESULTS: mRNAs of the four reprogramming factors were transfected into GEFs, and were localized in nucleus with approximately 90% transfection efficiency...
February 13, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28192181/in-vitro-development-of-zebrafish-vascular-networks
#3
Muhammad Ibrahim, Michael K Richardson
A major limitation to culturing tissues and organs is the lack of a functional vascular network in vitro. The zebrafish possess many useful properties which makes it a promising model for such studies. Unfortunately, methods of culturing endothelial cells from this species are not well characterised. Here, we tried two methods (embryoid body culture and organ explants from transgenic zebrafish kdrl:GFP embryos) to develop in vitro vascular networks. In the kdrl:GFP line, endothelial cells expresses green fluorescent protein, which allows to track the vascular development in live cultures...
February 9, 2017: Reproductive Toxicology
https://www.readbyqxmd.com/read/28191760/directing-differentiation-of-pluripotent-stem-cells-toward-retinal-pigment-epithelium-lineage
#4
Parul Choudhary, Heather Booth, Alex Gutteridge, Beata Surmacz, Irene Louca, Juliette Steer, Julie Kerby, Paul John Whiting
Development of efficient and reproducible conditions for directed differentiation of pluripotent stem cells into specific cell types is important not only to understand early human development but also to enable more practical applications, such as in vitro disease modeling, drug discovery, and cell therapies. The differentiation of stem cells to retinal pigment epithelium (RPE) in particular holds promise as a source of cells for therapeutic replacement in age-related macular degeneration. Here we show development of an efficient method for deriving homogeneous RPE populations in a period of 45 days using an adherent, monolayer system and defined xeno-free media and matrices...
February 2017: Stem Cells Translational Medicine
https://www.readbyqxmd.com/read/28182681/a-standardized-human-embryoid-body-platform-for-the-detection-and-analysis-of-teratogens
#5
Anthony Flamier, Supriya Singh, Theodore P Rasmussen
Teratogens are compounds that can induce birth defects upon exposure of the developing fetus. To date, most teratogen studies utilize pregnant rodents to determine compound teratogenicity in vivo. However, this is a low throughput approach that cannot easily meet the need for comprehensive high-volume teratogen assessment, a goal of the US Environmental Protection Agency. In addition, rodent and human development differ substantially, and therefore the use of assays using relevant human cells has utility. For these reasons, interest has recently focused on the use of human embryonic stem cells for teratogen assessment...
2017: PloS One
https://www.readbyqxmd.com/read/28163723/activin-a-modulates-cripto-1-hnf4%C3%AE-cells-to-guide-cardiac-differentiation-from-human-embryonic-stem-cells
#6
Robin Duelen, Guillaume Gilbert, Abdulsamie Patel, Nathalie de Schaetzen, Liesbeth De Waele, Llewelyn Roderick, Karin R Sipido, Catherine M Verfaillie, Gunnar M Buyse, Lieven Thorrez, Maurilio Sampaolesi
The use of human pluripotent stem cells in basic and translational cardiac research requires efficient differentiation protocols towards cardiomyocytes. In vitro differentiation yields heterogeneous populations of ventricular-, atrial-, and nodal-like cells hindering their potential applications in regenerative therapies. We described the effect of the growth factor Activin A during early human embryonic stem cell fate determination in cardiac differentiation. Addition of high levels of Activin A during embryoid body cardiac differentiation augmented the generation of endoderm derivatives, which in turn promoted cardiomyocyte differentiation...
2017: Stem Cells International
https://www.readbyqxmd.com/read/28159716/cellular-self-assembly-and-biomaterials-based-organoid-models-of-development-and-diseases
#7
REVIEW
Shivem B Shah, Ankur Singh
: Organogenesis and morphogenesis have informed our understanding of physiology, pathophysiology, and avenues to create new curative and regenerative therapies. Thus far, this understanding has been hindered by the lack of a physiologically relevant yet accessible model that affords biological control. Recently, three-dimensional ex vivo cellular cultures created through cellular self-assembly under natural extracellular matrix cues or through biomaterial-based directed assembly have been shown to physically resemble and recapture some functionality of target organs...
January 31, 2017: Acta Biomaterialia
https://www.readbyqxmd.com/read/28147316/generation-of-in-vivo-neural-stem-cells-using-partially-reprogrammed-cells-defective-in-in-vitro-differentiation-potential
#8
Jong Soo Kim, Yean Ju Hong, Hyun Woo Choi, Hyuk Song, Sung June Byun, Jeong Tae Do
Pluripotent stem cells can be easily differentiated in vitro into a certain lineage through embryoid body formation. Recently, however, we reported partially reprogrammed cells showing some pluripotent characteristics, which failed to differentiate in vitro. Here, we attempted to generate neural stem cells (NSCs) from partially reprogrammed cells using an in vivo differentiation system involving teratoma formation. Partially reprogrammed cells formed teratomas after injection into immunocompromised mice, and NSCs could be isolated from these teratomas...
January 27, 2017: Oncotarget
https://www.readbyqxmd.com/read/28123351/involvement-of-rictor-mtorc2-in-cardiomyocyte-differentiation-of-mouse-embryonic-stem-cells-in-vitro
#9
Bei Zheng, Jiadan Wang, Leilei Tang, Chao Tan, Zhe Zhao, Yi Xiao, Renshan Ge, Danyan Zhu
Rictor is a key regulatory/structural subunit of the mammalian target of rapamycin complex 2 (mTORC2) and is required for phosphorylation of Akt at serine 473. It plays an important role in cell survival, actin cytoskeleton organization and other processes in embryogenesis. However, the role of Rictor/mTORC2 in the embryonic cardiac differentiation has been uncovered. In the present study, we examined a possible link between Rictor expression and cardiomyocyte differentiation of the mouse embryonic stem (mES) cells...
2017: International Journal of Biological Sciences
https://www.readbyqxmd.com/read/28117672/reprogramming-methods-do-not-affect-gene-expression-profile-of-human-induced-pluripotent-stem-cells
#10
Marta Trevisan, Giovanna Desole, Giulia Costanzi, Enrico Lavezzo, Giorgio Palù, Luisa Barzon
Induced pluripotent stem cells (iPSCs) are pluripotent cells derived from adult somatic cells. After the pioneering work by Yamanaka, who first generated iPSCs by retroviral transduction of four reprogramming factors, several alternative methods to obtain iPSCs have been developed in order to increase the yield and safety of the process. However, the question remains open on whether the different reprogramming methods can influence the pluripotency features of the derived lines. In this study, three different strategies, based on retroviral vectors, episomal vectors, and Sendai virus vectors, were applied to derive iPSCs from human fibroblasts...
January 20, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/28106276/crispr-cas9-mediated-deletion-of-c1eis-inhibits-chicken-embryonic-stem-cell-differentiation-into-male-germ-cells-gallus-gallus
#11
Qisheng Zuo, Kai Jin, Yingjie Wang, Jiuzhou Song, Yani Zhang, Bichun Li
We previously found that C1EIS is preferentially expressed in Chicken spermatogonial stem cells (SSCs) by RNA sequencing (RNA-seq), so our current study focused on C1EIS's role in Chicken embryonic stem cells (ESCs) differentiation into male germ cells. We constructed a CRISPR/Cas9 vector targeting C1EIS. T7 endonuclease I (T7EI) digestion method and sequencing of TA cloning were used to detect the knock-out efficiency of the Single guide RNA (sgRNA) after the cas9/gRNA vector transfected into D fibroblasts 1(DF-1), ESCs and Chicken embryos...
January 20, 2017: Journal of Cellular Biochemistry
https://www.readbyqxmd.com/read/28068081/peptide-decorated-nanofibrous-niche-augments-in-vitro-directed-osteogenic-conversion-of-human-pluripotent-stem-cells
#12
Yi Deng, Yuanyi Yang, Shicheng Wei
Realization of clinical potential of human pluripotent stem cells (hPSCs) in bone regenerative medicine requires development of simple and safe biomaterials for expansion of hPSCs followed by directing their lineage commitment to osteoblasts. In the present study, a chemically defined peptide-decorated polycaprolactone (PCL) nanofibrous microenvironment was prepared through electrospinning technology and subsequent conjugation with vitronectin peptide to promote the culture and osteogenic potential of hPSCs in vitro...
January 24, 2017: Biomacromolecules
https://www.readbyqxmd.com/read/28066542/characterization-of-tubular-liquid-crystal-structure-in-embryonic-stem-cell-derived-embryoid-bodies
#13
MengMeng Xu, Odell D Jones, Liyang Wang, Xin Zhou, Harry G Davis, Joseph L Bryant, Jianjie Ma, Willian B Isaacs, Xuehong Xu
BACKGROUND: Massive liquid crystal droplets have been found during embryonic development in more than twenty different tissues and organs, including the liver, brain and kidney. Liquid crystal deposits have also been identified in multiple human pathologies, including vascular disease, liver dysfunction, age-related macular degeneration, and other chronic illnesses. Despite the involvement of liquid crystals in such a large number of human processes, this phenomenon is poorly understood and there are no in vitro systems to further examine the function of liquid crystals in biology...
2017: Cell & Bioscience
https://www.readbyqxmd.com/read/28017376/myogenic-potential-of-mouse-embryonic-stem-cells-lacking-functional-pax7-tested-in-vitro-by-5-azacitidine-treatment-and-in-vivo-in-regenerating-skeletal-muscle
#14
Anita Helinska, Maciej Krupa, Karolina Archacka, Areta M Czerwinska, Wladyslawa Streminska, Katarzyna Janczyk-Ilach, Maria A Ciemerych, Iwona Grabowska
Regeneration of skeletal muscle relies on the presence of satellite cells. Satellite cells deficiency accompanying some degenerative diseases is the reason for the search for the "replacement cells" that can be used in the muscle therapies. Due to their unique properties embryonic stem cells (ESCs), as well as myogenic cells derived from them, are considered as a promising source of therapeutic cells. Among the factors crucial for the specification of myogenic precursor cells is Pax7 that sustains proper function of satellite cells...
January 2017: European Journal of Cell Biology
https://www.readbyqxmd.com/read/28000905/teratomas-produced-from-human-pluripotent-stem-cells-xenografted-into-immunodeficient-mice-a-histopathology-atlas
#15
Ivan Damjanov, Peter W Andrews
This atlas illustrates the microscopic features of tumors produced from human pluripotent stem cells (hPSCs) xenografted into immunosuppressed mice, according to the generally accepted protocols for performing this teratoma assay of stem cell pluripotency. Microphotographs depict various hematoxylin and eosin (H&E) stained tissues derived from all three embryonic germ layers (ectoderm, mesoderm and endoderm). The appearance of persistent hPSC in teratomas is also described with special emphasis on the morphogenesis of embryoid bodies and yolk sac components surrounding them...
2016: International Journal of Developmental Biology
https://www.readbyqxmd.com/read/27987073/comparison-of-four-protocols-to-generate-chondrocyte-like-cells-from-human-induced-pluripotent-stem-cells-hipscs
#16
Wiktoria Maria Suchorska, Ewelina Augustyniak, Magdalena Richter, Tomasz Trzeciak
Stem cells (SCs) are a promising approach to regenerative medicine, with the potential to treat numerous orthopedic disorders, including osteo-degenerative diseases. The development of human-induced pluripotent stem cells (hiPSCs) has increased the potential of SCs for new treatments. However, current methods of differentiating hiPSCs into chondrocyte-like cells are suboptimal and better methods are needed. The aim of the present study was to assess four different chondrogenic differentiation protocols to identify the most efficient method of generating hiPSC-derived chondrocytes...
December 16, 2016: Stem Cell Reviews
https://www.readbyqxmd.com/read/27934689/differentiating-mouse-embryonic-stem-cells-into-embryoid-bodies-by-hanging-drop-cultures
#17
Richard Behringer, Marina Gertsenstein, Kristina Vintersten Nagy, Andras Nagy
Embryonic stem (ES) cells can develop into many types of differentiated tissues if they are placed into a differentiating environment. This can occur in vivo when the ES cells are injected into or aggregated with an embryo, or in vitro if their culture conditions are modified to induce differentiation. There are an increasing number of differentiating culture conditions that can bias the differentiation of ES cells into desired cell types. Determining the mechanisms that control ES cell differentiation into therapeutically important cell types is a quickly growing area of research...
December 1, 2016: Cold Spring Harbor Protocols
https://www.readbyqxmd.com/read/27934594/establishment-of-induced-pluripotent-stem-cell-ipsc-line-from-55-year-old-male-patient-with-hemorrhagic-moyamoya-disease
#18
Marina Cardano, Fabio Marsoner, Matteo Marcatili, Thodoris Karnavas, Jacopo Zasso, Luigi Andrea Lanterna, Luciano Conti
Peripheral blood mononuclear cells (PBMCs) were collected from 55-year old male patient with a confirmed diagnosis of hemorrhagic Moyamoya disease (MMD). PBMCs were reprogrammed using Sendai virus particles delivering the four Yamanaka factors. A footprint-free hiPSC line was characterized by the expression of pluripotency markers and a normal karyotype. These cells were able to give rise to Embryoid Bodies and to a progeny of differentiated cells belonging to the 3 germ layers. This hiPSC line represents a suitable tool for modelling in vitro MMD disease to investigate the cellular mechanisms underlying the occurrence of this pathology...
November 9, 2016: Stem Cell Research
https://www.readbyqxmd.com/read/27911036/properties-of-embryoid-bodies
#19
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
#20
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
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