Titration of differentiation medium components

摘要

Methods and composition for differentiation of pluripotent stem cells are provided. For example, in certain aspects methods including screening of optimal differentiation conditions for a selected stem cell clone in a selected batch of culture medium and use of the determined optimal condition for differentiation into a specific cell lineage.

主权项

1. A method for differentiating human pluripotent stem cells from a human pluripotent stem cell clone into a chosen cell lineage, comprising the steps of:
a) obtaining a human pluripotent stem cell clone prepared from an individual human pluripotent stem cell, the human stem cell clone being either a human embryonic stem cell clone or a human induced pluripotent stem cell clone; b) obtaining a batch of culture media; c) culturing human pluripotent stem cells from the pluripotent stem cell clone in a maintenance culture media prepared in a first amount of said batch of culture media, wherein the maintenance culture media maintains the human pluripotent stem cells in an undifferentiated stated without the presence of a fibroblast feed cell layer; d) culturing human pluripotent stem cells in a second amount of said batch of culture media in the presence of one or more differentiation factors capable of differentiating the human pluripotent stem cells into said chosen cell lineage, thereby determining amounts of one or more differentiation factors required to differentiate the human pluripotent stem cells into said chosen cell lineage; e) obtaining a differentiation media comprising the amounts of one or more differentiation factors required to differentiate the human pluripotent stem cells into the chosen cell lineage, as determined in step d; and f) differentiating human pluripotent stem cells in the differentiation medium of step e, thereby differentiating human pluripotent stem cells from a human pluripotent stem cell clone into a chosen cell lineage.