Method for preparing clonogenic fibroblasts, method for gene...

C - Chemistry – Metallurgy – 12 – N

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C12N 5/10 (2006.01) A61K 35/12 (2006.01) A61K 48/00 (2006.01) A61L 27/00 (2006.01) C07K 14/535 (2006.01) A61K 38/00 (2006.01) C12N 5/06 (2006.01)

Patent

CA 2183438

The invention relates to a process for preparing clonogenic fibroblasts, with tissue being removed from the donor and the individual cells being isolated from the tissue, the resulting cell suspension being strained, the cells which are contained in the cell suspension being washed and the cells being converted into a tissue culture, with the exception of the isolation of individual cells by mechanical comminution, followed by an enzymic treatment with collagenase alone, and with at least one gene being inserted into the fibroblasts by means of the transfection, which gene encodes a biologically active protein, preferably a therapeutically active protein, for example a growth factor, a hormone, an enzyme, a coagulation factor or a coagulation inhibitor.

Dans la méthode proposée pour préparer des fibroplasts clonogènes, on prélève un tissu sur un donneur et l'on en isole des cellules; la suspension cellulaire ainsi obtenue est passée au tamis; les cellules contenues dans la suspension sont lavées et transférée en culture tissulaire; une possibilité consiste à isoler les cellules par fragmentation mécanique et à les soumettre ensuite à un traitement enzymatique avec de la collagénase uniquement. La transfection dans les fibroblasts entraîne l'injection d'au moins un gène codant pour une protéine biologiquement active, de préférence une protéine active du point de vue thérapeutique, par exemple un facteur de croissance, une hormone, une enzyme, un facteur ou un inhibiteur de coagulation.

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