Abstract：
:Reliable and efficient DNA and RNA transfection methods are required when studying the role of individual genes in mouse pluripotent stem cells. However, these cells usually grow in tight clusters and are therefore more difficult to transfect than many other cell lines. We have found that transfection is especially challenging when mouse embryonic stem (mES) cells are cultured in the newly described 2i medium, which is based on two chemical inhibitors of differentiation pathways. In the present study we have performed a side-by-side comparison of commercially available, non-viral transfection reagents with regard to their ability to deliver plasmid DNA and siRNA into adherent and/or trypsinized mES cells cultured in 2i medium, assessing transfection rates, plasmid gene expression, siRNA mediated knockdown of Oct4 and viability. Finally, we present a fast and efficient method for transfection of trypsinized mES cells using the liposomal-based Lipofectamine 2000. With only a five-minute long transfection time we obtained at least 85 % transfected cells with 80 % maintained viability. Moreover, this protocol saves up to a day of experimental time since the cells are in suspension at the time of transfection, which allows for immediately re-plating into the appropriate format. This fast, simplified and highly efficient transfection method will be valuable for both basic research and high-throughput applications.
journal_name
Stem Cell Rev Repjournal_title
Stem cell reviews and reportsauthors
Tamm C,Kadekar S,Pijuan-Galitó S,Annerén Cdoi
10.1007/s12015-016-9673-5subject
Has Abstractpub_date
2016-10-01 00:00:00pages
584-591issue
5eissn
2629-3269issn
2629-3277pii
10.1007/s12015-016-9673-5journal_volume
12pub_type
杂志文章abstract：:Mesenchymal stromal cells (MSCs) constitute a heterogeneous population of stromal cells with immunomodulatory and regenerative properties that support their therapeutic use. MSCs isolated from many tissue sources replicate vigorously in vitro and maintain their main biological properties allowing their widespread clin...
journal_title：Stem cell reviews and reports
pub_type： 杂志文章,评审
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abstract：:Zinc-finger nucleases (ZFNs) are sequence-specific genome engineering tools with great potential for the development of gene therapies. The achievement of permanent cures through gene therapy requires targeting of stem cells but the effects and/or side effects of ZFN treatment on adult stem cell potency are largely un...
journal_title：Stem cell reviews and reports
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abstract：:The discovery of induced pluripotent stem (iPS) cells in 2006 marked a major breakthrough in regenerative medicine, enabling reversal of terminally differentiated somatic cells into pluripotent stem cells. The embryonic stem (ES) cells-like pluripotency and unlimited self-renewal capability of iPS cells have granted t...
journal_title：Stem cell reviews and reports
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journal_title：Stem cell reviews and reports
pub_type： 杂志文章
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更新日期：2014-04-01 00:00:00
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journal_title：Stem cell reviews and reports
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abstract：:Neural progenitor cells (NPCs) are multipotent cells that have the potential to produce neurons and glial cells in the neural system. NPCs undergo identity maintenance or differentiation regulated by different kinds of transcription factors. Here we present evidence that ETV5, which is an ETS transcription factor, pro...
journal_title：Stem cell reviews and reports
pub_type： 杂志文章
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journal_title：Stem cell reviews and reports
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journal_title：Stem cell reviews and reports
pub_type： 杂志文章,评审
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更新日期：2011-06-01 00:00:00
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journal_title：Stem cell reviews and reports
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journal_title：Stem cell reviews and reports
pub_type： 杂志文章,评审
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更新日期：2014-12-01 00:00:00
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journal_title：Stem cell reviews and reports
pub_type： 杂志文章,评审
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更新日期：2014-06-01 00:00:00
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journal_title：Stem cell reviews and reports
pub_type： 杂志文章
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journal_title：Stem cell reviews and reports
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journal_title：Stem cell reviews and reports
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更新日期：2015-02-01 00:00:00
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journal_title：Stem cell reviews and reports
pub_type： 杂志文章,已发布勘误
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更新日期：2020-06-01 00:00:00
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journal_title：Stem cell reviews and reports
pub_type： 杂志文章,评审
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更新日期：2012-12-01 00:00:00
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journal_title：Stem cell reviews and reports
pub_type： 杂志文章,评审
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更新日期：2009-06-01 00:00:00
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journal_title：Stem cell reviews and reports
pub_type： 杂志文章,评审
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journal_title：Stem cell reviews and reports
pub_type： 杂志文章,评审
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journal_title：Stem cell reviews and reports
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journal_title：Stem cell reviews and reports
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journal_title：Stem cell reviews and reports
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journal_title：Stem cell reviews and reports
pub_type： 杂志文章
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更新日期：2011-09-01 00:00:00
abstract：:The use of mesenchymal stem/stromal cells (MSC) in regenerative medicine often requires MSC to function in environments of high oxidative stress. Human pregnancy is a condition where the mother's tissues, and in particular her circulatory system, are exposed to increased levels of oxidative stress. MSC in the maternal...
journal_title：Stem cell reviews and reports
pub_type： 杂志文章
doi：10.1007/s12015-016-9649-5
更新日期：2016-06-01 00:00:00
abstract：BACKGROUND:Mesenchymal stem cells derived from the chorionic villi of human term placenta (pMSCs) have drawn considerable interest because of their multipotent differentiation potential and their immunomodulatory capacity. These properties are the foundation for their clinical application in the fields of stem cell tra...
journal_title：Stem cell reviews and reports
pub_type： 杂志文章
doi：10.1007/s12015-014-9562-8
更新日期：2015-06-01 00:00:00
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journal_title：Stem cell reviews and reports
pub_type： 杂志文章,评审
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更新日期：2017-06-01 00:00:00
abstract：:Endothelial progenitors are a population of cells with the inherent capacity to differentiate into mature endothelial cells and proangiogenic paracrine action. These characteristics have led to extensive studies being performed and tested in the treatment of tissue ischemia. The natural course of diabetes mellitus (DM...
journal_title：Stem cell reviews and reports
pub_type： 杂志文章,评审
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更新日期：2019-04-01 00:00:00
abstract：:The intestine integrates the function of digestion, absorption, and barrier, which is easily damaged by the external factors upon ageing. The intestinal stem cells (ISCs) exist at the intestinal crypt base and play an indispensable role in intestinal homeostasis and regeneration. The intestine ageing contributes to ma...
journal_title：Stem cell reviews and reports
pub_type： 杂志文章,评审
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更新日期：2020-11-17 00:00:00
abstract：:Human embryonic stem cells (hESCs) are especially resistant to several cellular stresses, but the existence and induction of Endoplasmic Reticulum (ER) stress by culture conditions are unknown. Using qPCR, here, we investigated the behavior of the principal sensors of ER stress and their relation with the feeder layer...
journal_title：Stem cell reviews and reports
pub_type： 杂志文章
doi：10.1007/s12015-010-9135-4
更新日期：2010-09-01 00:00:00
abstract：:The interest in the use of stem cells as a source for therapy has increased dramatically over the last decades. Different stem cell types have been tested in both in vitro and in vivo models, because of their properties such as differentiation potential, trophic effects and immune modulatory properties. To further opt...
journal_title：Stem cell reviews and reports
pub_type： 杂志文章
doi：10.1007/s12015-014-9575-3
更新日期：2015-04-01 00:00:00
