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Method Article
Pascale V Guillot
Guillot PV Lab
Corresponding Author
License:
This work is licensed under a CC BY-NC 3.0 License. Read Full License
Pluripotent stem cells have potential applications in regenerative medicine, disease modelling and drug screening. Induced pluripotent stem (iPS) cells have first been generated from fibroblasts using retroviral insertion of OCT4A, SOX2, c-MYC and KLF4. Since then, a number of methods have been developed to avoid the random integration of ectopic factors in the genome and the low efficiency of the process. Those include alternative integrating, non-integrating, excisable and DNA-free systems, but they all present challenges that prevail their use as a clinical and molecular tool. Here we present a transgene-free detailed protocol to generate human pluripotent cells from c-KIT+ amniotic fluid fetal stem cells. The parental populations express OCT4A and can be reverted to functional pluripotency through manipulations of culture conditions and Valproic acid (VPA) supplementation. The resulting pluripotent cells could potentially be used safely without ethical and legal restriction in the clinic for prenatal and postnatal autologous use.
Keywords
reprogramming, stem cells, amniotic fluid, pluripotency, valproic acid
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Associated Publications
Valproic Acid Confers Functional Pluripotency to Human Amniotic Fluid Stem Cells in a Transgene-free Approach
by Dafni Moschidou, Sayandip Mukherjee, Michael P Blundell, +13
Molecular Therapy (03 July, 2012)
An open repository of community-contributed protocols sponsored by Nature Research.
Learn more about Protocol Exchange
Method Article
Pascale V Guillot
Guillot PV Lab
Corresponding Author
Version History
CURRENT STATUS: Posted
Version 1
Posted 06 Jul, 2012
No community comments so far
Metrics
Comments: 0
HTML Views: ...
License:
This work is licensed under a CC BY-NC 3.0 License. Read Full License
Pluripotent stem cells have potential applications in regenerative medicine, disease modelling and drug screening. Induced pluripotent stem (iPS) cells have first been generated from fibroblasts using retroviral insertion of OCT4A, SOX2, c-MYC and KLF4. Since then, a number of methods have been developed to avoid the random integration of ectopic factors in the genome and the low efficiency of the process. Those include alternative integrating, non-integrating, excisable and DNA-free systems, but they all present challenges that prevail their use as a clinical and molecular tool. Here we present a transgene-free detailed protocol to generate human pluripotent cells from c-KIT+ amniotic fluid fetal stem cells. The parental populations express OCT4A and can be reverted to functional pluripotency through manipulations of culture conditions and Valproic acid (VPA) supplementation. The resulting pluripotent cells could potentially be used safely without ethical and legal restriction in the clinic for prenatal and postnatal autologous use.
Figure 1
None
This is a list of supplementary files associated with this preprint. Click to download.
- supplement0.doc
Table 1 Troubleshooting table
Loading...
Associated Publications
Valproic Acid Confers Functional Pluripotency to Human Amniotic Fluid Stem Cells in a Transgene-free Approach
by Dafni Moschidou, Sayandip Mukherjee, Michael P Blundell, +13
Molecular Therapy (03 July, 2012)