Methods for neural differentiation of embryonic stem cells...

C - Chemistry – Metallurgy – 12 – N

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C12N 5/00 (2006.01) A01N 1/02 (2006.01) C12N 15/85 (2006.01) C12N 15/86 (2006.01)

Patent

CA 2521039

The present invention provides methods for human pluripotent cell culturing and for neural cell production. More particularly, the present invention provides culturing methods employing dissociating cell cultures to an essentially single cell culture, such as by employing antibody selection and bulk passaging treatments utilizing the subsequent application of Collagenase and trypsin. In certain embodiments, the cells are further treated with essentially serum free MEDII conditioned medium, proline, or minimal medium, and are optionally treated with amphiphilic lipid compounds for the generation of human neural cells from pluripotent human cells. In certain embodiments, the cells cultured using these methods have an abnormal karyotype.

L'invention concerne des procédés de culture de cellules pluripotentes humaines et de production de cellules neuronales. Plus particulièrement, l'invention concerne des procédés de culture consistant à dissocier des cultures cellulaires de manière à obtenir une culture cellulaire sensiblement unique, par exemple par sélection d'anticorps et par des traitements de passage en masse par application ultérieure de collagénase et de trypsine. Dans certains modes de réalisation, les cellules sont également traitées au moyen d'un milieu conditionné MEDII sensiblement dépourvu de sérum, de proline, ou d'un milieu minimum, et sont éventuellement traitées au moyen de composés lipidiques amphiphiles afin que soient générées des cellules neuronales humaines à partir de cellules pluripotentes humaines. Dans certains modes de réalisation, les cellules cultivées au moyen de ces procédés présentent un caryotype anormal.

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