Cryopreservation of human blastocyst-derived stem cells by...

A - Human Necessities – 01 – N

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A01N 1/02 (2006.01) C12N 5/0735 (2010.01)

Patent

CA 2524929

An improved method for vitrification of biological cells, especially blastocyst-derived stem cells (BS cells). The method is very mild for the cells that remain viable after they have been thawed. The method comprises, i) transfer of the cells to a first solution (solution A), ii) optionally incubation of the cells in the first solution, iii) transfer the cells obtained in step i) or ii) to a second solution (solution B), iv) optionally incubation of the cells in the second solution, v) transfer of the cells obtained from step iii) or iv) into one or more closed straws with dimensions that allow a volume of at least 20 ~l to be contained in them vi) sealing the one or more closed straws, and vii) vitrification of the one or more closed straws. An important feature of the present invention is the use of closed straw and that relatively large volumes can be efficiently vitrified and subsequently thawed.

L'invention concerne un procédé amélioré de vitrification de cellules biologiques, plus spécifiquement de cellules souches dérivées de blastocystes. Ce procédé est très doux pour les cellules qui restent viables après avoir été décongelées. Ce procédé comprend i) le transfert des cellules dans une première solution (solution A), ii) éventuellement l'incubation des cellules dans la première solution, iii) le transfert des cellules obtenues à l'étape i) ou ii) dans une seconde solution (solution B), iv) éventuellement l'incubation des cellules dans la seconde solution, v) le transfert des cellules obtenues à l'étape iii) ou iv) dans une ou plusieurs paillettes fermées dont les dimensions permettent qu'un volume d'au moins 20 ?l soit contenu dans ces paillettes, vi) le scellement hermétique de cette ou de ces paillettes fermées et vii) la vitrification de la ou des paillettes fermées. Une importante caractéristique de cette invention est l'utilisation de paillettes fermées ainsi que le fait que des volumes relativement grands peuvent être efficacement vitrifiés puis décongelés.

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