Method for enhancing keratinocyte stem cells

C - Chemistry – Metallurgy – 12 – N

Patent

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Details

C12N 5/071 (2010.01) A61K 35/36 (2006.01) A61L 27/60 (2006.01) A61K 35/12 (2006.01)

Patent

CA 2464196

The invention concerns a method for enriching a population of keratinocyte stem cells, from a preparation of keratinocytes, comprising a pre-enrichment step in keratinocyte stem cells by adherence of said preparation on a constituent of extracellular matrices, in particular recognized by molecules involved in the adherence of the keratinocytes, in particular by the integrin family receptors, particularly collagen, for a period of time between about 5 and about 30 minutes, preferably between about 10 and about 20 minutes, advantageously 12 minutes, and in recuperating the stem cells which have adhered on said constituent of extracellular matrices by separation in conditions enabling to maintain the viabilty of the stem cells which have adhered, in particular by trypsinization, particularly using trypsin 0.05 % and EDTA 0.02 % to obtain a population of adherent cells.

L'invention concerne un procédé d'enrichissement d'une population de cellules souches kératinocytaires, à partir d'une préparation de kératinocytes, comprenant une étape de pré-enrichissement en cellules souches kératinocytaires par adhésion de ladite préparation sur un composant de matrices extracellulaires, notamment reconnu par des molécules impliquées dans l'adhésion des kératinocytes, notamment par des récepteurs de la famille des intégrines, notamment du collagène, pendant une durée d'environ 5 mn à environ 30 mn, notamment d'environ 10 mn à environ 20 mn, avantageusement 12 mn, et la récupération des cellules souches ayant adhéré sur le susdit composant de matrices extracellulaires par décollement dans des conditions permettant de maintenir la viabilité des cellules souches ayant adhéré, notamment par trypsination, notamment à l'aide de trypsine 0,05 % et EDTA 0,02 %, pour obtenir une population de cellules adhérentes.

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