Genome-wide targets identification of “core” pluripotency transcription factors with integrated features in human embryonic stem cells

2016 ◽  
Vol 12 (4) ◽  
pp. 1324-1332 ◽  
Author(s):  
Leijie Li ◽  
Zhaobin Chen ◽  
Liangcai Zhang ◽  
Guiyou Liu ◽  
Jinlian Hua ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Transcription Factors ◽  
Embryonic Stem Cells ◽  
Embryonic Stem Cell ◽  
Human Embryonic Stem Cells ◽  
Human Embryonic Stem Cell ◽  
Embryonic Stem ◽  
Genome Wide ◽  
Novel Model

LMA: A novel model to predict target of pluripotency transcriptional factors in human embryonic stem cell.

scholarly journals Human embryonic stem cells: towards therapies for cardiac disease. Derivation of a Dutch human embryonic stem cell line

2005 ◽  
Vol 11 (4) ◽  
pp. 476-485 ◽  
Author(s):  
Anja van de Stolpe ◽  
Stieneke van den Brink ◽  
Marga van Rooijen ◽  
Dorien Ward-van Oostwaard ◽  
Wouter van Inzen ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Embryonic Stem Cells ◽  
Embryonic Stem Cell ◽  
Human Embryonic Stem Cells ◽  
Cardiac Disease ◽  
Human Embryonic Stem Cell ◽  
Embryonic Stem Cell Line ◽  
Embryonic Stem ◽  
Stem Cell Line

P–797 A novel method for establishing human embryonic stem cells independent of feeder cells

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
B Cai
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Embryonic Stem Cells ◽  
Embryonic Stem Cell ◽  
Success Rate ◽  
Cell Lines ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Feeder Cells ◽  
Novel Method

Abstract Study question Is there a efficient establishing method of human embryonic stem cells directly from the human blastocysts independent of feeder cells? Summary answer We established a novel method of generating human embryonic stem cells directly from human blastocysts independent of feeder layer cells. What is known already Establishing embryonic stem cells lines mainly needed to coculture ICM clumps with feeder cells (like mouse or human fibroblasts) ,this brought in potential heterogeneous pollution.Although there had be some reports about generating human ESCs independent of feeder cells,but the efficiency was low and conditioned medium were unstable and also had the biological contamination. Study design, size, duration We used ten day5/6 donated human blastocysts from our reproductive center ,most of them were genetically diseased embryos with abnormal PGT diagnosis.After establishing ESCs procedure , all the cell lines were identified with pluripotency and differentiation potential tests.The success rate of system was calculated and compared with the conventional methods. Participants/materials, setting, methods In brief, ICM clumps were separated mechanically by using a micromanipulation system,and then transferred to a 30ul mTESR plus culture media drop pretreated with the geltrex (1:100 dilution) matrix and oxygen concentration was 5%. When cells attached and migrated,we also used laser to destroy the remaining trophoblast cells.About 10 days,the typical ES clone can be mechanically passaged and cells can be cultured in normal oxygen concentrations after passage 2. . Main results and the role of chance Using this method we had successfully established nine embryonic stem cell lines from donated human blastocysts ,the success rate was 90% (9/10). Each cell lines had passed the evaluation test of embryonic stem cell. When compared with the conventional feeder cells dependent method,our novol methods not only eliminated the pollution from heterogeneous cells,but also had higher success rate (90% vs 25%). Limitations, reasons for caution Due to the scarcity of donated human blastocysts, this experiment was a single-center experiment with small samples. Wider implications of the findings: We speculated that the batch differences of culture dishes, matrix and culture medium might affect the establish efficiency , and how to carry out a high level of quality control work might be the key factor to keep the system stable. Trial registration number basic research

P-797 A novel method for establishing human embryonic stem cells independent of feeder cells

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
B Cai
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Embryonic Stem Cells ◽  
Embryonic Stem Cell ◽  
Success Rate ◽  
Cell Lines ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Feeder Cells ◽  
Novel Method

Abstract Study question Is there a efficient establishing method of human embryonic stem cells directly from the human blastocysts independent of feeder cells? Summary answer We established a novel method of generating human embryonic stem cells directly from human blastocysts independent of feeder layer cells. What is known already Establishing embryonic stem cells lines mainly needed to coculture ICM clumps with feeder cells (like mouse or human fibroblasts), this brought in potential heterogeneous pollution. Although there had be some reports about generating human ESCs independent of feeder cells, but the efficiency was low and conditioned medium were unstable and also had the biological contamination. Study design, size, duration We used ten day5/6 donated human blastocysts from our reproductive center, most of them were genetically diseased embryos with abnormal PGT diagnosis. After establishing ESCs procedure, all the cell lines were identified with pluripotency and differentiation potential tests. The success rate of system was calculated and compared with the conventional methods. Participants/materials, setting, methods In brief, ICM clumps were separated mechanically by using a micromanipulation system,and then transferred to a 30ul mTESR plus culture media drop pretreated with the geltrex (1:100 dilution) matrix and oxygen concentration was 5%. When cells attached and migrated,we also used laser to destroy the remaining trophoblast cells. About 10 days,the typical ES clone can be mechanically passaged and cells can be cultured in normal oxygen concentrations after passage 2.. Main results and the role of chance Using this method we had successfully established nine embryonic stem cell lines from donated human blastocysts, the success rate was 90% (9/10). Each cell lines had passed the evaluation test of embryonic stem cell. When compared with the conventional feeder cells dependent method,our novol methods not only eliminated the pollution from heterogeneous cells,but also had higher success rate (90% vs 25%). Limitations, reasons for caution Due to the scarcity of donated human blastocysts, this experiment was a single-center experiment with small samples. Wider implications of the findings We speculated that the batch differences of culture dishes, matrix and culture medium might affect the establish efficiency, and how to carry out a high level of quality control work might be the key factor to keep the system stable. Trial registration number basic research

scholarly journals Use of human embryonic stem cells to model pediatric gliomas with H3.3K27M histone mutation

Science ◽  
2014 ◽  
Vol 346 (6216) ◽  
pp. 1529-1533 ◽  
Author(s):  
Kosuke Funato ◽  
Tamara Major ◽  
Peter W. Lewis ◽  
C. David Allis ◽  
Viviane Tabar
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Neural Progenitor Cells ◽  
Embryonic Stem ◽  
Cell System ◽  
Genome Wide ◽  
Regulator Genes

Over 70% of diffuse intrinsic pediatric gliomas, an aggressive brainstem tumor, harbor heterozygous mutations that create a K27M amino acid substitution (methionine replaces lysine 27) in the tail of histone H3.3. The role of the H3.3K27M mutation in tumorigenesis is not fully understood. Here, we use a human embryonic stem cell system to model this tumor. We show that H3.3K27M expression synergizes with p53 loss and PDGFRA activation in neural progenitor cells derived from human embryonic stem cells, resulting in neoplastic transformation. Genome-wide analyses indicate a resetting of the transformed precursors to a developmentally more primitive stem cell state, with evidence of major modifications of histone marks at several master regulator genes. Drug screening assays identified a compound targeting the protein menin as an inhibitor of tumor cell growth in vitro and in mice.

scholarly journals Similar Population of CD133+ and DDX4+ VSEL-Like Stem Cells Sorted from Human Embryonic Stem Cell, Ovarian, and Ovarian Cancer Ascites Cell Cultures: The Real Embryonic Stem Cells?

Cells ◽  
2019 ◽  
Vol 8 (7) ◽  
pp. 706 ◽  
Author(s):  
Irma Virant-Klun ◽  
Petra Skerl ◽  
Srdjan Novakovic ◽  
Eda Vrtacnik-Bokal ◽  
Spela Smrkolj
Keyword(s):  
Ovarian Cancer ◽  
Stem Cells ◽  
Stem Cell ◽  
Embryonic Stem Cells ◽  
Embryonic Stem Cell ◽  
Cell Cultures ◽  
Human Embryonic Stem Cell ◽  
Embryonic Stem ◽  
Recurrent Ovarian Cancer

A population of small stem cells with diameters of up to 5 μm resembling very small embryonic-like stem cells (VSELs) were sorted from human embryonic stem cell (hESC) cultures using magnetic-activated cell sorting (MACS) based on the expression of a stem-cell-related marker prominin-1 (CD133). These VSEL-like stem cells had nuclei that almost filled the whole cell volume and expressed stem-cell-related markers (CD133, SSEA-4) and markers of germinal lineage (DDX4/VASA, PRDM14). They were comparable to similar populations of small stem cells sorted from cell cultures of normal ovaries and were the predominant cells in ascites of recurrent ovarian cancer. The sorted populations of CD133+ VSEL-like stem cells were quiescent in vitro, except for ascites, and were highly activated after exposure to valproic acid and follicle-stimulating hormone (FSH), indicating a new tool to study these cells in vitro. These VSEL-like stem cells spontaneously formed clusters resembling tumour-like structures or grew into larger, oocyte-like cells and were differentiated in vitro into adipogenic, osteogenic and neural lineages after sorting. We propose the population of VSEL-like stem cells from hESC cultures as potential original embryonic stem cells, which are present in the human embryo, persist in adult human ovaries from the embryonic period of life and are involved in cancer manifestation.

scholarly journals Embryonic stem cells: protein interaction networks

2011 ◽  
Vol 2 (1-2) ◽  
pp. 13-25 ◽  
Author(s):  
Patricia Miang-Lon Ng ◽  
Thomas Lufkin
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Transcription Factors ◽  
Embryonic Stem Cells ◽  
Embryonic Stem Cell ◽  
Protein Interaction ◽  
Protein Interaction Network ◽  
Embryonic Stem ◽  
Interaction Network ◽  
Cell Protein

AbstractEmbryonic stem cells have the ability to differentiate into nearly all cell types. However, the molecular mechanism of its pluripotency is still unclear. Oct3/4, Sox2 and Nanog are important factors of pluripotency. Oct3/4 (hereafter referred to as Oct4), in particular, has been an irreplaceable factor in the induction of pluripotency in adult cells. Proteins interacting with Oct4 and Nanog have been identified via affinity purification and mass spectrometry. These data, together with iterative purifications of interacting proteins allowed a protein interaction network to be constructed. The network currently includes 77 transcription factors, all of which are interconnected in one network. In-depth studies of some of these transcription factors show that they all recruit the NuRD complex. Hence, transcription factor clustering and chromosomal remodeling are key mechanism used by embryonic stem cells. Studies using RNA interference suggest that more pluripotency genes are yet to be discovered via protein-protein interactions. More work is required to complete and curate the embryonic stem cell protein interaction network. Analysis of a saturated protein interaction network by system biology tools can greatly aid in the understanding of the embryonic stem cell pluripotency network.

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