scholarly journals Efficient Generation of Hepatoblasts From Human ES Cells and iPS Cells by Transient Overexpression of Homeobox Gene HEX

2011 ◽  
Vol 19 (2) ◽  
pp. 400-407 ◽  
Author(s):  
Mitsuru Inamura ◽  
Kenji Kawabata ◽  
Kazuo Takayama ◽  
Katsuhisa Tashiro ◽  
Fuminori Sakurai ◽  
...  
Keyword(s):  
Es Cells ◽  
Homeobox Gene ◽  
Ips Cells ◽  
Efficient Generation ◽  
Human Es Cells ◽  
Transient Overexpression

scholarly journals Highly efficient differentiation of human ES cells and iPS cells into mature pancreatic insulin-producing cells

Cell Research ◽  
2009 ◽  
Vol 19 (4) ◽  
pp. 429-438 ◽  
Author(s):  
Donghui Zhang ◽  
Wei Jiang ◽  
Meng Liu ◽  
Xin Sui ◽  
Xiaolei Yin ◽  
...  
Keyword(s):  
Es Cells ◽  
Ips Cells ◽  
Highly Efficient ◽  
Insulin Producing Cells ◽  
Pancreatic Insulin ◽  
Human Es Cells

Efficient Production of Human Hematopoietic Progenitors from Human Pluripotent Stem Cells Using Chemically Defined Media without Serum or Feeder Cells

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 2463-2463
Author(s):  
Zhaohui Ye ◽  
Xiaobing Yu ◽  
Linzhao Cheng
Keyword(s):  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Es Cells ◽  
Ips Cells ◽  
Feeder Cells ◽  
Hematopoietic Differentiation ◽  
Hematopoietic Progenitors ◽  
Serum Factors ◽  
Defined Media ◽  
Human Es Cells

Abstract As established cell lines, human pluripotent stem cells such as embryonic stem (ES) or induced pluripotent stem (iPS) cells can divide indefinitely while retaining their potential to differentiate into many cell types in culture. The induction of mesoderm formation and hematopoietic differentiation was achieved either via embryoid body (EB) formation by culturing ES cell aggregation in suspension or by co-cultures with mouse stromal cell lines. Traditionally serum factors are also added for mesoderm induction and hematopoietic differentiation. Although hematopoietic progenitors are obtained from multiple lines of human ES cells, the low efficiency and high variability have hindered the progress of using human ES cells as a model for studying human hematopoiesis. Normally <15% of cells obtained from a primary culture expressed CD34 (a marker for endothelial cells and hematopoietic progenitors) and even less for CD45 (a pan-leukocyte marker). To generate maximal output of CD34+CD45+ hematopoietic progenitors, we decided to adopt the serum-free and spin-EB formation method (Ng, Blood, 2005) and systematically improved culture conditions. 3,000 human ES cells were added into each well in 96-well plates and formed an aggregate after centrifugation. BMP4 and bFGF were added at day 1, and VEGF and hematopoietic cytokines was added at day 3–9. VEGF was then withdrawn after day 9. Single EB (occasionally 2) grew in each well. By day 8, small blast (or lymphocyte-) like cells were observed on the edge of EBs. By day 12–14, we observed the outgrowth of blast cells (in hundreds to thousands) surrounding each EB (Panel A). By FACS analysis (Panel B), we observed nearly 50% of the total cells express CD45 at day 12, and many co-express CD34. The lymphocytelike cells can be easily separately from EBs by passing through a 40-micron strainer, and nearly all the isolated cells express CD45 (and 50–75% of them co-express CD34). We obtained 6 million CD45+ cells from 0.9 million human ES cells 14 days after EB formation. We also observed that the conditional HES1-ER transgene expression further increased the frequency of CD34+CD45+ cells as we observed under a different culture condition (Yu. Cell Stem Cells, 2008). The isolated CD45+ cells formed efficiently hematopoietic colonies in the methylcellulose medium with a frequency of ~58+/− 4 colonies per 3,000 cells, with or without the HES1-ER transgene. We are currently testing in vivo activities of isolated CD45+ cell populations (+/− HES1-ER) in the NOD/SCID/γC−/− mice. We are also testing if this improved and defined method would also be applicable to hematopoietic differentiation of human iPS cells we recently derived (Mali, Stem Cells, 2008). The significantly improved method using defined media in the absence of serum factors or feeder cells warrants further investigation whether it is better and more reproducible to elucidate mechanisms that regulate early human hematopoiesis, and to generate a large quantity of CD34+CD45+ human hematopoietic progenitor cells for various applications. Figure Figure

scholarly journals Matrix Metalloproteinase-3 in Odontoblastic Cells Derived from Ips Cells: Unique Proliferation Response as Odontoblastic Cells Derived from ES Cells

PLoS ONE ◽  
2013 ◽  
Vol 8 (12) ◽  
pp. e83563 ◽  
Author(s):  
Taiki Hiyama ◽  
Nobuaki Ozeki ◽  
Makio Mogi ◽  
Hideyuki Yamaguchi ◽  
Rie Kawai ◽  
...  
Keyword(s):  
Matrix Metalloproteinase ◽  
Es Cells ◽  
Ips Cells ◽  
Matrix Metalloproteinase 3 ◽  
Proliferation Response

scholarly journals The chromosomal protein SMCHD1 regulates DNA methylation and the 2c-like state of embryonic stem cells by antagonizing TET proteins

2021 ◽  
Vol 7 (4) ◽  
pp. eabb9149
Author(s):  
Zhijun Huang ◽  
Jiyoung Yu ◽  
Wei Cui ◽  
Benjamin K. Johnson ◽  
Kyunggon Kim ◽  
...  
Keyword(s):  
Es Cells ◽  
Embryonic Stem ◽  
Homeobox Gene ◽  
Dna Demethylation ◽  
Chromosomal Protein ◽  
Dna Hypomethylation ◽  
Tet Proteins ◽  
Target Sites ◽  
Structural Maintenance Of Chromosomes ◽  
Hinge Domain

5-Methylcytosine (5mC) oxidases, the ten-eleven translocation (TET) proteins, initiate DNA demethylation, but it is unclear how 5mC oxidation is regulated. We show that the protein SMCHD1 (structural maintenance of chromosomes flexible hinge domain containing 1) is found in complexes with TET proteins and negatively regulates TET activities. Removal of SMCHD1 from mouse embryonic stem (ES) cells induces DNA hypomethylation, preferentially at SMCHD1 target sites and accumulation of 5-hydroxymethylcytosine (5hmC), along with promoter demethylation and activation of the Dux double-homeobox gene. In the absence of SMCHD1, ES cells acquire a two-cell (2c) embryo–like state characterized by activation of an early embryonic transcriptome that is substantially imposed by Dux. Using Smchd1/Tet1/Tet2/Tet3 quadruple-knockout cells, we show that DNA demethylation, activation of Dux, and other genes upon SMCHD1 loss depend on TET proteins. These data identify SMCHD1 as an antagonist of the 2c-like state of ES cells and of TET-mediated DNA demethylation.

scholarly journals The Effects of Tgfb1 and Csf3 on Chondrogenic Differentiation of iPS Cells in 2D and 3D Culture Environment

2021 ◽  
Vol 22 (6) ◽  
pp. 2978
Author(s):  
Chie-Hong Wang ◽  
Chun-Hao Tsai ◽  
Tsung-Li Lin ◽  
Shih-Ping Liu
Keyword(s):  
Cell Proliferation ◽  
Chondrogenic Differentiation ◽  
Expression Profiles ◽  
Es Cells ◽  
3D Culture ◽  
Ips Cells ◽  
3D Cultures ◽  
Positive Effects ◽  
Cartilaginous Matrix ◽  
Culture Environment

Mesenchymal stem (MS) cells, embryonic stem (ES) cells, and induced pluripotent stem (iPS) cells are known for their ability to differentiate into different lineages, including chondrocytes in culture. However, the existing protocol for chondrocyte differentiation is time consuming and labor intensive. To improve and simplify the differentiation strategy, we have explored the effects of interactions between growth factors (transforming growth factor β1 (Tgfb1) and colony stimulating factor 3 (Csf3), and culture environments (2D monolayer and 3D nanofiber scaffold) on chondrogenic differentiation. For this, we have examined cell morphologies, proliferation rates, viability, and gene expression profiles, and characterized the cartilaginous matrix formed in the chondrogenic cultures under different treatment regimens. Our data show that 3D cultures support higher proliferation rate than the 2D cultures. Tgfb1 promotes cell proliferation and viability in both types of culture, whereas Csf3 shows positive effects only in 3D cultures. Interestingly, our results indicate that the combined treatments of Tgfb1 and Csf3 do not affect cell proliferation and viability. The expression of cartilaginous matrix in different treatment groups indicates the presence of chondrocytes. We found that, at the end of differentiation stage 1, pluripotent markers were downregulated, while the mesodermal marker was upregulated. However, the expression of chondrogenic markers (col2a1 and aggrecan) was upregulated only in the 3D cultures. Here, we report an efficient, scalable, and convenient protocol for chondrogenic differentiation of iPS cells, and our data suggest that a 3D culture environment, combined with tgfb1 and csf3 treatment, promotes the chondrogenic differentiation.

scholarly journals Utilization of the AAVS1 safe harbor locus for hematopoietic specific transgene expression and gene knockdown in human ES cells

2014 ◽  
Vol 12 (3) ◽  
pp. 630-637 ◽  
Author(s):  
Amita Tiyaboonchai ◽  
Helen Mac ◽  
Razveen Shamsedeen ◽  
Jason A. Mills ◽  
Siddarth Kishore ◽  
...  
Keyword(s):  
Transgene Expression ◽  
Es Cells ◽  
Gene Knockdown ◽  
Safe Harbor ◽  
Human Es Cells

Physiological low oxygen during development induces neural differentiation from mouse ES cells and mouse iPS cells

2010 ◽  
Vol 68 ◽  
pp. e244
Author(s):  
Masataka Fujita ◽  
Tea-Sun Kim ◽  
Sachiyo Misumi ◽  
Yoshitomo Ueda ◽  
Hitoo Nishino ◽  
...  
Keyword(s):  
Neural Differentiation ◽  
Es Cells ◽  
Ips Cells ◽  
Low Oxygen ◽  
Mouse Es Cells

Neural Differentiation of Human ES Cells

2007 ◽  
Vol 36 (1) ◽  
Author(s):  
Malkiel A. Cohen ◽  
Pavey Itsykson ◽  
Benjamin E. Reubinoff
Keyword(s):  
Neural Differentiation ◽  
Es Cells ◽  
Human Es Cells

Human embryonic stem cells

2000 ◽  
Vol 113 (1) ◽  
pp. 5-10 ◽  
Author(s):  
M.F. Pera ◽  
B. Reubinoff ◽  
A. Trounson
Keyword(s):  
Stem Cells ◽  
Cell Lines ◽  
Embryonal Carcinoma ◽  
Es Cells ◽  
Embryonic Stem ◽  
Early Embryo ◽  
Carcinoma Cells ◽  
Mouse Es Cells ◽  
Undifferentiated State ◽  
Human Es Cells

Embryonic stem (ES) cells are cells derived from the early embryo that can be propagated indefinitely in the primitive undifferentiated state while remaining pluripotent; they share these properties with embryonic germ (EG) cells. Candidate ES and EG cell lines from the human blastocyst and embryonic gonad can differentiate into multiple types of somatic cell. The phenotype of the blastocyst-derived cell lines is very similar to that of monkey ES cells and pluripotent human embryonal carcinoma cells, but differs from that of mouse ES cells or the human germ-cell-derived stem cells. Although our understanding of the control of growth and differentiation of human ES cells is quite limited, it is clear that the development of these cell lines will have a widespread impact on biomedical research.

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