scholarly journals Replication Initiation Patterns in the β-Globin Loci of Totipotent and Differentiated Murine Cells: Evidence for Multiple Initiation Regions

2002 ◽  
Vol 22 (2) ◽  
pp. 442-452 ◽  
Author(s):  
Mirit I. Aladjem ◽  
Luo Wei Rodewald ◽  
Chii Mai Lin ◽  
Sarah Bowman ◽  
Daniel M. Cimbora ◽  
...  
Keyword(s):  
Stem Cells ◽  
Dna Replication ◽  
Embryonic Stem Cells ◽  
Chromatin Structure ◽  
Embryonic Stem ◽  
Replication Initiation ◽  
Globin Genes ◽  
Primary Sequence ◽  
Differentiated Cells ◽  
Hypersensitive Sites

ABSTRACT The replication initiation pattern of the murine β-globin locus was analyzed in totipotent embryonic stem cells and in differentiated cell lines. Initiation events in the murine β-globin locus were detected in a region extending from the embryonic Ey gene to the adult βminor gene, unlike the restricted initiation observed in the human locus. Totipotent and differentiated cells exhibited similar initiation patterns. Deletion of the region between the adult globin genes did not prevent initiation in the remainder of the locus, suggesting that the potential to initiate DNA replication was not contained exclusively within the primary sequence of the deleted region. In addition, a deletion encompassing the six identified 5′ hypersensitive sites in the mouse locus control region had no effect on initiation from within the locus. As this deletion also did not affect the chromatin structure of the locus, we propose that the sequences determining both chromatin structure and replication initiation lie outside the hypersensitive sites removed by the deletion.

scholarly journals Epigenome and chromatin structure in human embryonic stem cells undergoing differentiation

2008 ◽  
Vol 237 (12) ◽  
pp. 3690-3702 ◽  
Author(s):  
Eva Bártová ◽  
Gabriela Galiová ◽  
Jana Krejčí ◽  
Andrea Harničarová ◽  
Luděk Strašák ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Chromatin Structure ◽  
Embryonic Stem

Long targeting arms do not increase the efficiency of homologous recombination in the β-globin locus of murine embryonic stem cells

Blood ◽  
2003 ◽  
Vol 102 (4) ◽  
pp. 1531-1533 ◽  
Author(s):  
Zhi Hong Lu ◽  
Jason T. Books ◽  
Richard M. Kaufman ◽  
Timothy J. Ley
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Homologous Recombination ◽  
Globin Gene ◽  
Embryonic Stem ◽  
Regulatory Sequences ◽  
Globin Genes ◽  
Gene Correction ◽  
Murine Embryonic Stem Cells ◽  
Alternative Approaches

Abstract The correction of mutant β-globin genes has long been a therapeutic goal for patients with β-thalassemia or hemoglobinopathies. The use of homologous recombination (HR) to achieve this goal is an attractive approach because it eliminates the need to include regulatory sequences in the therapeutic construct, and it eliminates mutagenesis induced by random integration. However, HR is a very inefficient process for gene correction, and its efficiency is probably locus dependent. The length of targeting arms is thought to be a determinant of targeting efficiency, so we compared the ability of standard (8-kb) versus very long (16-, 24-, and 110-kb) regions of homology to correct a mutant murine β-globin gene in embryonic stem cells. Increasing the length of the targeting sequences did not increase the efficiency of HR in this locus, suggesting that alternative approaches will be required to improve the efficiency of this approach for globin gene correction.

scholarly journals The Molecular Basis for the Lack of Inflammatory Responses in Mouse Embryonic Stem Cells and Their Differentiated Cells

2017 ◽  
Vol 198 (5) ◽  
pp. 2147-2155 ◽  
Author(s):  
William D’Angelo ◽  
Chandan Gurung ◽  
Dhiraj Acharya ◽  
Bohan Chen ◽  
Natalya Ortolano ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Molecular Basis ◽  
Inflammatory Responses ◽  
Embryonic Stem ◽  
Mouse Embryonic Stem Cells ◽  
Differentiated Cells

scholarly journals Identification of novel proteins differentially expressed in pluripotent embryonic stem cells and differentiated cells

2015 ◽  
Vol 62 (3.4) ◽  
pp. 130-136 ◽  
Author(s):  
Kei Enomoto ◽  
Kanako Watanabe-Susaki ◽  
Megumi Kowno ◽  
Hitomi Takada ◽  
Atsushi Intoh ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Differentially Expressed ◽  
Novel Proteins ◽  
Differentiated Cells

scholarly journals A Bivalent Chromatin Structure Marks Key Developmental Genes in Embryonic Stem Cells

Cell ◽  
2006 ◽  
Vol 125 (2) ◽  
pp. 315-326 ◽  
Author(s):  
Bradley E. Bernstein ◽  
Tarjei S. Mikkelsen ◽  
Xiaohui Xie ◽  
Michael Kamal ◽  
Dana J. Huebert ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Chromatin Structure ◽  
Embryonic Stem ◽  
Developmental Genes

scholarly journals Defining totipotency using criteria of increasing stringency

2020 ◽  
Author(s):  
Eszter Posfai ◽  
John Paul Schell ◽  
Adrian Janiszewski ◽  
Isidora Rovic ◽  
Alexander Murray ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Single Cell ◽  
Pluripotent Stem Cells ◽  
Embryonic Stem ◽  
Cell Types ◽  
Differentiated Cells ◽  
Totipotent Cell

AbstractTotipotency is the ability of a single cell to give rise to all the differentiated cells that build the conceptus, yet how to capture this property in vitro remains incompletely understood. Defining totipotency relies upon a variety of assays of variable stringency. Here we describe criteria to define totipotency. We illustrate how distinct criteria of increasing stringency can be used to judge totipotency by evaluating candidate totipotent cell types in the mouse, including early blastomeres and expanded or extended pluripotent stem cells. Our data challenge the notion that expanded or extended pluripotent states harbor increased totipotent potential relative to conventional embryonic stem cells under in vivo conditions.

scholarly journals Differentiation of Human Adipose-Derived Stem Cells into Parathyroid Hormone-Secreting Cells

2020 ◽  
Author(s):  
Eun Heui Kim ◽  
Sang Soo Kim ◽  
Ju In Kim ◽  
Ji Min Cheon ◽  
Joo Hyoung Kim ◽  
...  
Keyword(s):  
Stem Cells ◽  
Vitamin D ◽  
Parathyroid Hormone ◽  
Embryonic Stem Cells ◽  
Parathyroid Gland ◽  
Embryonic Stem ◽  
Thymic Epithelial Cells ◽  
Adipose Derived Stem Cells ◽  
Adipose Tissues ◽  
Differentiated Cells

Abstract IntroductionComplications arising from neck surgery are the most common causes of acquired hypoparathyroidism. Parathyroid hormone (PTH) regulates serum calcium and vitamin D levels. The current treatments for hypoparathyroidism, including PTH supplements and high doses of calcium and vitamin D, are a life-long burden and may cause side effects. Recent studies have shown that parathyroid gland differentiation occurs from embryonic stem cells, thymic epithelial cells, and tonsil-derived stem cells. Because adipose-derived stem cells (ADSCs) do not face ethical concerns raised for studies with human embryonic stem cells, in the present study, we attempted to induce differentiation of ADSCs into parathyroid cells.MethodsADSCs were isolated from abdominal subcutaneous adipose tissues obtained by plastic surgery from three male donors (mean age, 40 years). For differentiation into parathyroid-like cells, ADSCs were incubated in minimum essential medium-alpha supplemented with activin A and soluble Sonic hedgehog for 7–21 d. The parathyroid differentiation markers PTH, glial cells missing homolog 2 (GCM2), and chemokine (C-C motif) ligand 21 (CCL21) were detected using real-time quantitative polymerase chain reaction to confirm differentiation.ResultsThe ADSCs exhibited flat and vacuolated morphology, which changed into secretory parathyroid gland–like nodules on day 21 after differentiation. The mRNA expression of PTH, GCM2, and CCL21 increased in the differentiated cells. Furthermore, a significant amount of PTH protein was detected in differentiated cells on day 7 post-differentiation.ConclusionHuman ADSCs isolated from adipose tissues successfully differentiated into PTH-secreting cells. ADSC-secreted PTH may be a promising therapeutic for hypoparathyroidism patients.

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