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Method for producing high-quality iPS cells

  • US 10,544,396 B2
  • Filed: 07/11/2016
  • Issued: 01/28/2020
  • Est. Priority Date: 07/10/2015
  • Status: Active Grant
First Claim
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1. A method for producing an iPS cell in vitro, comprising:

  • i) introducing (a) a nuclear reprogramming substance, wherein the nuclear reprogramming substance consists of a polynucleotide encoding an Oct3/4 protein, a polynucleotide encoding a Sox2 protein, and a polynucleotide encoding a Klf4 protein, and (b) a polynucleotide encoding an H1foo protein into a somatic cell, andii) culturing the somatic cell from i) to produce an iPS cell, wherein the iPS cell, when compared to an iPS cell produced without introducing the polynucleotide encoding an H1foo protein, is improved as shown in one or more selected from the group consisting of the following (1) to (11);

    (1) an increase in a percentage of embryoid body formation when iPS cells have been cultured in a culture medium for embryoid body formation;

    (2) an increase in a number of embryoid body formation when iPS cells have been cultured in the culture medium for embryoid body formation;

    (3) a decrease in a size variance of embryoid bodies when iPS cells have been cultured in the culture medium for embryoid body formation;

    (4) an increase in a percentage of viable cells when iPS cells have been cultured in the culture medium for embryoid body formation;

    (5) a decrease in a percentage of apoptotic cells when iPS cells have been cultured in the culture medium for embryoid body formation;

    (6) an increase in an expression level of a Ki67 gene or a PCNA gene when iPS cells have been cultured in the culture medium for embryoid body formation;

    (7) an increase in a chimera competency of iPS cells;

    (8) an increase in a germline transmission potential of iPS cells;

    (9) an increase in an expression level of a SRF gene in iPS cells;

    (10) an increase in an expression level of a ACTG2 gene in iPS cells; and

    (11) a decrease in a variance of an expression level of a Oct3/4 gene in iPS cells.

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