Culture system for rapid expansion of human embryonic stem...

C - Chemistry – Metallurgy – 12 – N

Patent

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C12N 5/00 (2006.01) C12N 5/0735 (2010.01) C12N 5/10 (2006.01) C12N 15/00 (2006.01) C12N 15/87 (2006.01)

Patent

CA 2459957

This disclosure provides an improved system for culturing human pluripotent stem cells. Traditionally, pluripotent stem cells are cultured on a layer of feeder cells (such as mouse embryonic fibroblasts) to prevent them from differentiating. In the system described here, the role of feeder cells is replaced by components added to the culture environment that support rapid proliferation without differentiation. Effective features are a suitable support structure for the cells, and an effective medium that can be added fresh to the culture without being preconditioned by another cell type. Culturing human embryonic stem cells in fresh medium according to this invention causes the cells to expand surprisingly rapidly, while retaining the ability to differentiate into cells representing all three embryonic germ layers. This new culture system allows for bulk proliferation of pPS cells for commercial production of important products for use in drug screening and human therapy.

L'invention concerne un système amélioré de culture de cellules souches multipotentes humaines. On cultive généralement les cellules souches multipotentes sur une couche de cellules nourricières (p. ex. fibroblastes embryonnaires de souris) pour éviter qu'elles se différencient. Dans le système décrit, le rôle des cellules nourricières est remplacé par des constituants ajoutés au milieu de culture et qui permettent une prolifération rapide sans différenciation. Le système est caractérisé par une structure support appropriée pour les cellules ainsi qu'un milieu efficace pouvant être ajouté frais à la culture sans nécessiter de traitement préalable par un autre type cellulaire. La culture de cellules souches embryonnaires humaines en milieu frais, mise en oeuvre selon l'invention, permet d'obtenir une expansion étonnamment rapide des cellules et de maintenir la capacité de celles-ci à se différencier en cellules représentant les trois feuillets embryonnaires. Ce nouveau système de culture permet d'obtenir une prolifération globale de cellules pPS à des fins de production commerciale de produits importants pour le criblage de médicaments et la thérapie humaine.

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