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Reprogramming Normal and Cancerous Human Cells Lines into Human Induced Pluripotent Stem Cells - Research Paper Example

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Reprogramming Normal and Cancerous Human Cells Lines into Human Induced Pluripotent Stem Cells by Co-electroporation with Living Xenopus laevis Frog Oocytes Sergei Paylian Bioquark, Inc., P.O. Box 622, Lake Delton, WI 53940 USA Corresponding author: Tel.:+6084322227; fax: +6082530530, e-mail: info@bioquark.com Abstract Unique information on the ability of some amphibians to regenerate severely damaged tissues though the de-differentiation and re-differentiation of cells is encoded in the genetic makeup of amphibian oocytes…
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Reprogramming Normal and Cancerous Human Cells Lines into Human Induced Pluripotent Stem Cells
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Download file to see previous pages The iPSc-like cells generated by this study resemble human embryonic stem cells in colony morphology and expression of stem cell-associated transcription factors, including Oct3/4, Nanog, SOX-2, Rex-1, TRA-1-60, SSEA-1, and SSEA-4. The efficacy of reprogramming CD4+ lymphocytes into iPSc-like cells was 23.4 ± 3.5 %. Advantageously, this new method obviates the use of retroviral or lentiviral gene delivery vectors and other “non-parental” reprogramming approaches and may hold great promise as a strategy for the rapid and inexpensive production of human autologous stem cells. Keywords: human iPS cells, reprogramming, frog oocytes, Co-electroporation 1. ...
Recently, new, non-viral approaches for improving RP efficacy were reported. These methods include the use of recombinant proteins (Zhou et al., 2009); the use of DHP-derivative (novel anti-oxidant) and low oxygen-tension conditions (Jee et al., 2010). We believe that the low efficacy of reprogramming may be explained by the fact that the RP tools currently used in studies differ considerably from the delicate reprogramming machinery naturally present in living organisms. Reprogramming events observed in mammalian somatic cells induced by Xenopus laevis egg extracts were described by Miyamoto and colleagues (Miyamoto et al., 2007). At Bioquark, Inc., we developed a novel technique that allows the utilization of the natural reprogramming potential of living Xenopus laevis oocytes. We tried to generate human iPS cells in a consistent, safe and controllable way. Our system is fast, efficient, free of ethical controversy, highly reproducible, controllable, effects real time reprogramming, segregates human and amphibian components and simultaneously activates mutual semiochemical interactions and is able to produce partially reprogrammed cells which may represent the correct transitional point for successful re-differentiation or trans-differentiation reprogramming. This new experimental protocol utilizes the electroporation of living Xenopus laevis oocytes as a powerful RP tool through co-electroporating living Xenopus laevis oocytes with human donor cells in one electroporation chamber. We refer to this new procedure as “BQ-activation.” In our studies, we demonstrated that co-electroporation, with pulses of 150 v/cm / 25 µF / 7, of living Xenopus laevis oocytes ...Download file to see next pagesRead More
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