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Protein-induced pluripotent cell technology and uses thereof

  • US 9,816,076 B2
  • Filed: 05/02/2012
  • Issued: 11/14/2017
  • Est. Priority Date: 05/02/2011
  • Status: Active Grant
First Claim
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1. A method of generating human protein-induced pluripotent stem cells (piPSC), comprising:

  • i) producing reprogramming proteins by bacterial expression, wherein the reprogramming proteins comprise the combination of Klf-4, Oct-4, Sox-2, c-myc (KOSM);

    ii) modifying the bacterially expressed reprogramming proteins produced from step i) by mixing the proteins with a QQ-reagent comprising polyethylenimine (PEI) and DOTAP/DOPE;

    iii) delivering the modified reprogramming proteins produced from step ii) into nuclei of starting human somatic cells, using a QQ-protein transduction technique;

    iv) repeating several cell reprogramming cycles sufficient to create human reprogrammed piPSC;

    v) moving the human reprogrammed piPSC from step iv into a feeder-free medium for expansion; and

    vi) expanding and passaging the human reprogrammed piPSC from step v) as a whole dish of cells, with the proviso that colony picking is not performed, thereby generating homogeneous human piPSC.

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