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Derivation and culture of human embryo-derived cells

  • US 9,359,590 B2
  • Filed: 11/16/2005
  • Issued: 06/07/2016
  • Est. Priority Date: 11/16/2004
  • Status: Active Grant
First Claim
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1. A method of derivation of embryo-derived cells or embryo-derived cell lines consisting of the following steps:

  • a) culturing an embryo under low oxygen tension conditions of about 5% oxygen to the blastocyst stage;

    b) separating Inner Cell Mass (ICM) cells of said blastocyst from trophectodermal cells, plating said ICM cells and culturing said ICM cells under low oxygen tension conditions of about 5% oxygen for a time sufficient to produce embryonic cell-like outgrowths;

    c) at least once, dissociating said embryonic cell-like outgrowths, replating said dissociated embryonic cell-like outgrowths and culturing said dissociated embryonic cell-like outgrowths under low oxygen tension conditions of about 5% oxygen for a time sufficient to produce further embryonic cell-like outgrowths;

    d) analyzing said further embryonic cell-like outgrowths of step c) for embryonic stem cell-like properties;

    e) selecting outgrowths with embryonic stem cell-like properties; and

    f) isolating and dissociating said outgrowths with embryonic stem cell-like properties selected in step e) and culturing the dissociated cells under low oxygen tension conditions of about 5% oxygen to obtain embryo-derived cells or embryo-derived cell lines.

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