CPC C12Q 1/42 (2013.01) [G01N 33/5005 (2013.01); G01N 33/56966 (2013.01)] | 22 Claims |
1. A method of confirming absence of contaminating pluripotent stem cells in a preparation of differentiated cells generated by differentiation of said pluripotent stem cells, comprising:
(a) providing a preparation of differentiated cells produced by in vitro differentiation of pluripotent stem cells in a differentiation medium, and culturing said preparation of differentiated cells in a stem cell medium, wherein the stem cell medium maintains pluripotent stem cells in their pluripotent state;
(b) applying a first stain and a second stain to said preparation of differentiated cells, wherein
said first stain detects a first embryonic stem cell marker, expression of which is indicative of presence of said contaminating pluripotent stem cells, and
said second stain detects a second embryonic stem cell marker, expression of which is indicative of presence of said contaminating pluripotent stem cells,
wherein said first embryonic stem cell marker is different from said second embryonic stem cell marker,
wherein said first embryonic stem cell maker is alkaline phosphatase; and
(c) confirming absence of contaminating pluripotent stem cells, if no cells positive for both said first stain and said second stain are identified by microscopic observation.
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