Culturing human embryonic stem cells

C - Chemistry – Metallurgy – 12 – N

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C12N 5/0735 (2010.01) C12N 5/02 (2006.01)

Patent

CA 2579652

Previous methods for culturing human embryonic stem cells have required either fibroblast feeder cells or a medium, which has been exposed to fibroblast feeder cells in order to maintain the stem cells in an undifferentiated state. It has now been found that if high levels of fibroblast growth factor are used in a medium with gamma amino butyric acid, pipecholic acid, lithium and lipids, the stem cells will remain undifferentiated indefinitely through multiple passages, even without feeder cells or conditioned medium. A humanized matrix of human proteins can be used as a basement matrix to culture the cells. New lines of human embryonic stem cells made using these culture conditions, the medium and the matrix, will never have been exposed to animal cells, animal products, feeder cells or conditioned medium.

Les procédés précédents pour la culture de cellules souches embryonnaires humaines nécessitaient soit des cellules fibroblastes nourricières ou un milieu, qui a été exposé à des cellules fibroblastes nourricières afin de maintenir les cellules souches dans un état non différencié. On a maintenant découvert que si de niveaux élevés de facteur de croissance fibroblaste sont utilisés dans un milieu avec de l'acide gamma amino butyrique, de l'acide pipecolique, du lithium et des lipides, les cellules souches vont rester non différenciées indéfiniment à travers plusieurs passages, sans cellules nourricières ou milieu conditionné. Une matrice humanisée de cellules souches embryonnaires humaines peut être utilisée comme matrice de base pour la culture des cellules. De nouvelles lignées de cellules souches embryonnaires humaines fabriquées à l'aide de ces conditions de culture, le milieu et la matrice, n'auront jamais été exposés à des cellules animales, des produits d'origine animale, des cellules nourricières ou un milieu conditionné.

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