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Related Concept Videos

RNA-based Reprogramming of Human Primary Fibroblasts into Induced Pluripotent Stem Cells11:38

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Here we describe a clinically relevant, high-efficiency, feeder-free method to reprogram human primary fibroblasts into induced pluripotent stem cells using modified mRNAs encoding reprogramming factors and mature microRNA-367/302 mimics. Also included are methods to assess reprogramming efficiency, expand clonal iPSC colonies, and confirm expression of the pluripotency marker...
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The protocol presented in this study describes methods for the real-time monitoring of reprogramming progression via the kinetic measurement of positive and negative pluripotent stem cell markers using flow cytometry analysis. The protocol also includes the imaging-based assessment of morphology, and marker or reporter expression during iPSC...
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We demonstrate the protocol for the generation of induced pluripotent stem cells from human somatic cells using lentivirus-mediated delivery of the human factors Oct4, Sox2, Nanog, and Lin28. Pluripotency was confirmed by morphology and the presence of embryonic stem (ES) cell-specific...
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Kinetic Measurement and Real Time Visualization of Somatic Reprogramming
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Human oocytes reprogram somatic cells to a pluripotent state.

Scott Noggle1, Ho-Lim Fung, Athurva Gore

  • 1The New York Stem Cell Foundation Laboratory, New York, New York, USA.

Nature
|October 8, 2011
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Summary

Genome exchange in human oocytes for disease modeling failed due to developmental arrest. However, adding a somatic genome to an intact oocyte enabled blastocyst development and pluripotent stem cell derivation.

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Area of Science:

  • Reproductive biology
  • Stem cell science
  • Human genetics

Background:

  • Cell replacement therapy holds promise for treating degenerative human diseases.
  • Generating patient-specific cells requires reprogramming somatic cells into pluripotent stem cells.

Purpose of the Study:

  • To investigate the feasibility of generating patient-specific pluripotent stem cells via human oocyte genome exchange.
  • To identify factors limiting successful reprogramming and development.

Main Methods:

  • Human oocytes underwent genome exchange with somatic cell nuclei.
  • Alternatively, somatic cell nuclei were added to intact oocytes, creating triploid cells.
  • Derived cells were assessed for developmental potential and pluripotency markers.

Main Results:

  • Genome exchange led to developmental arrest at late cleavage stages with transcriptional abnormalities.
  • Addition of somatic genomes to intact oocytes resulted in blastocyst development.
  • Stem cell lines derived from these blastocysts exhibited pluripotency and differentiated into all three germ layers.

Conclusions:

  • Removal of the oocyte's genome is the primary cause of developmental failure in genome-exchanged oocytes.
  • Reprogramming human somatic cells using oocytes is feasible when the oocyte genome is retained.
  • This approach enables the generation of patient-specific pluripotent stem cells for disease modeling and potential therapies.