Cationic star copolymers based on β-cyclodextrins for efficient gene delivery to mouse embryonic stem cell colonies

2015 ◽  
Vol 51 (54) ◽  
pp. 10815-10818 ◽  
Author(s):  
Xian Jun Loh ◽  
Yun-Long Wu
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Embryonic Stem Cells ◽  
Gene Transfer ◽  
Embryonic Stem ◽  
Mouse Embryonic Stem Cell ◽  
Nonviral Gene Transfer ◽  
Star Copolymers ◽  
Efficient Gene ◽  
Star Copolymer

A cationic star copolymer with a β-cyclodextrin core was developed for nonviral gene transfer to mouse embryonic stem cells (mESCs).

scholarly journals Epsins Regulate Mouse Embryonic Stem Cell Exit from Pluripotency and Neural Commitment by Controlling Notch Activation

2019 ◽  
Vol 2019 ◽  
pp. 1-13 ◽  
Author(s):  
Marina Cardano ◽  
Jacopo Zasso ◽  
Luca Ruggiero ◽  
Giuseppina Di Giacomo ◽  
Matteo Marcatili ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Embryonic Stem Cells ◽  
Notch Signaling ◽  
Neural Differentiation ◽  
Radial Glia ◽  
Embryonic Stem ◽  
Mouse Embryonic Stem Cell ◽  
Neural Progenitors ◽  
Notch Activation

Epsins are part of the internalization machinery pivotal to control clathrin-mediated endocytosis. Here, we report that epsin family members are expressed in mouse embryonic stem cells (mESCs) and that epsin1/2 knockdown alters both mESC exits from pluripotency and their differentiation. Furthermore, we show that epsin1/2 knockdown compromises the correct polarization and division of mESC-derived neural progenitors and their conversion into expandable radial glia-like neural stem cells. Finally, we provide evidence that Notch signaling is impaired following epsin1/2 knockdown and that experimental restoration of Notch signaling rescues the epsin-mediated phenotypes. We conclude that epsins contribute to control mESC exit from pluripotency and allow their neural differentiation by appropriate modulation of Notch signaling.

Efficient Transduction of Common Marmoset (Callithrix jacchus) Hematopoietic and Embryonic Stem Cells Using Foamyvirus Vectors.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 5530-5530
Author(s):  
Peter A. Horn ◽  
Melanie Wurm ◽  
Ryo Kurita ◽  
Tomoko Yokoo ◽  
Rainer Blasczyk ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Gene Transfer ◽  
Nonhuman Primate ◽  
Embryonic Stem ◽  
Common Marmoset ◽  
Transduction Efficiency ◽  
Nonhuman Primate Model ◽  
Primate Model ◽  
Efficient Gene

Abstract Preclinical animal models are important for evaluating the safety and therapeutic efficacy of new therapeutic modalities such as gene therapy. From the different large animal models, nonhuman primate models have emerged over the last decades as highly desirable experimental systems from both a pathophysiologic and pharmacokinetic viewpoint and the study of nonhuman primates has provided important information on the efficacy and safety of gene therapy systems in vivo prior to human trials. The common marmoset (Callithrix jacchus) has the advantage that it is a small, and thus relatively inexpensive nonhuman primate model. Currently, very little data on the transduction efficiency of foamyviral vectors for gene transfer into marmoset stem cells exists. We therefore performed a direct comparison using identically designed gammaretroviral, lentiviral and foamyviral vector constructs expressing the enhanced green fluorescent protein (EGFP) from the spleen focus forming virus (SFFV) promoter pseudotyped with either the modified human foamy virus (HFV) envelope EM140 or the G-protein of vesicular stomatitis virus (VSV-G) for the transduction of common marmoset embryonic stem cells (CMES) as well as marmoset CD34+ hematopoietic progenitor cells. Virus stocks of these vectors were prepared by polyethyleneimine-mediated transfection of 293T cells and concentrated approximately 10-fold by centrifugation for 4 hours at 10.000 g at 4°C. Three different target cell populations were transduced: common marmoset embryonic stem cells (CMES) or cryopreserved CD34-enriched cells from bone marrow of a common marmoset either after a two-day prestimulation in the presence of IL-6, FLT3L, cSCF and TPO at a concentration of 100 ng/mL each, or after overnight incubation with 100 ng/mL SCF only. Equal numbers of cells were exposed to the four different vector preparations for 14 hours in 12-well dishes coated with CH-296. The read-out was based on fluorescence microscopy of colonies plated in methyl cellulose as well as flow cytometry (FACS). Foamyviral vectors with the foamyviral envelope were the most efficient gene transfer tool for marmoset hematopoietic CD34-positive cells with stable transduction rates of over 80% as assessed by flow cytometry at both 2 or 7 days after the end of transduction and on average 88% transduction efficiency into colony forming cells (CFU-C). Transduction of CFU-C with all other vector preparations was below 60%. In CMES, initial gene transfer rates of over 80% were achieved with the VSV-G pseudotype lentiviral vector, however, expression decreased to 13% after 7 days. In contrast, the foamyviral vector pseudotyped with the foamyviral envelope decreased only from 49% to 24% after 7 days. In conclusion, we achieved stable viral gene transfer and expression in CMES cells as well as highly efficient gene transfer into common marmoset hematopoietic CD34 positive cells using foamyviral vectors. These results suggest that foamyviral vectors may be highly feasible vectors for stem cell gene transfer and thus set the stage for a more detailed analysis of this vector system in transplantation studies in this nonhuman primate model.

Efficient gene transfer of a simian immuno-deficiency viral vector into cardiomyocytes derived from primate embryonic stem cells

2003 ◽  
Vol 5 (11) ◽  
pp. 921-928 ◽  
Author(s):  
Mihoko Nagata ◽  
Masafumi Takahashi ◽  
Shin-ichi Muramatsu ◽  
Yasuji Ueda ◽  
Yutaka Hanazono ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Gene Transfer ◽  
Viral Vector ◽  
Embryonic Stem ◽  
Efficient Gene

scholarly journals Highly Efficient Gene Transfer into Primate Embryonic Stem Cells with a Simian Lentivirus Vector

2002 ◽  
Vol 6 (2) ◽  
pp. 162-168 ◽  
Author(s):  
Takayuki Asano ◽  
Yutaka Hanazono ◽  
Yasuji Ueda ◽  
Shin-ichi Muramatsu ◽  
Akihiro Kume ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Gene Transfer ◽  
Embryonic Stem ◽  
Lentivirus Vector ◽  
Highly Efficient ◽  
Efficient Gene

scholarly journals Efficient gene transfer into mouse embryonic stem cells with adenovirus vectors

2005 ◽  
Vol 12 (3) ◽  
pp. 547-554 ◽  
Author(s):  
Kenji Kawabata ◽  
Fuminori Sakurai ◽  
Teruhide Yamaguchi ◽  
Takao Hayakawa ◽  
Hiroyuki Mizuguchi
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Gene Transfer ◽  
Embryonic Stem ◽  
Mouse Embryonic Stem Cells ◽  
Efficient Gene ◽  
Adenovirus Vectors

scholarly journals Microcircuit formation following transplantation of mouse embryonic stem cell-derived neurons in peripheral nerve

2017 ◽  
Vol 117 (4) ◽  
pp. 1683-1689 ◽  
Author(s):  
Philippe Magown ◽  
Victor F. Rafuse ◽  
Robert M. Brownstone
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Embryonic Stem Cells ◽  
Embryonic Stem Cell ◽  
Spontaneous Activity ◽  
Self Assembly ◽  
Embryonic Stem ◽  
Mouse Embryonic Stem Cell ◽  
Muscle Contractions ◽  
Functional Connections

Motoneurons derived from embryonic stem cells can be transplanted in the tibial nerve, where they extend axons to functionally innervate target muscle. Here, we studied spontaneous muscle contractions in these grafts 3 mo following transplantation. One-half of the transplanted grafts generated rhythmic muscle contractions of variable patterns, either spontaneously or in response to brief electrical stimulation. Activity generated by transplanted embryonic stem cell-derived neurons was driven by glutamate and was modulated by muscarinic and GABAergic/glycinergic transmission. Furthermore, rhythmicity was promoted by the same transmitter combination that evokes rhythmic locomotor activity in spinal cord circuits. These results demonstrate that there is a degree of self-assembly of microcircuits in these peripheral grafts involving embryonic stem cell-derived motoneurons and interneurons. Such spontaneous activity is reminiscent of embryonic circuit development in which spontaneous activity is essential for proper connectivity and function and may be necessary for the grafts to form functional connections with muscle. NEW & NOTEWORTHY This manuscript demonstrates that, following peripheral transplantation of neurons derived from embryonic stem cells, the grafts are spontaneously active. The activity is produced and modulated by a number of transmitter systems, indicating that there is a degree of self-assembly of circuits in the grafts.

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