Authors
Dafni Moschidou and Pascale V Guillot
Summary
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.Further details
The protocol was published on Protocol Exchange on 6 July 2012. To see the entire protocol, click on the source link.Advertisement
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