C - Chemistry – Metallurgy – 12 – N
Patent
C - Chemistry, Metallurgy
12
N
C12N 5/02 (2006.01) A61K 35/14 (2006.01) C12M 1/12 (2006.01) C12M 3/00 (2006.01) A61K 35/12 (2006.01) C12N 5/06 (2006.01)
Patent
CA 2194270
Disclosed is a method for cultivating macrophages from blood, including collecting a quantity of blood, separating from the quantity of blood, a white blood cell fraction that includes red blood cells and white blood Cells including monocytes, the separating including fractionating the quantity of blood, by centrifuging the quantity of blood in a centrifuge container, into a plasma fraction and the white blood Cell fraction and segregating the white blood cell faction from the plasma fraction, differentiating the monocytes into macrophages by an osmotic shock to the white blood cell fraction, the osmotic shock being more destructive of red blood cells than white blood cells and lysing at least some of the red blood cells and some, but not all, of the white blood cells and reestablishing isotonicity in the white blood cell fraction, by introducing a hypertonic solution into the white blood Cell fraction thereby creating an isotonic solution of the white blood cell fraction, the isotonic solution being characterized by a saline concentration.
L'invention concerne un procédé de mise en culture de cellules présentes dans une suspension qui consiste à placer les cellules, un ou plusieurs réactifs et un milieu de culture dans des récipients stériles séparés reliés entre eux et fermés hermétiquement par des tubulures stériles dotées de dispositifs de régulation du débit de fluide et pouvant être ouverts et fermés manuellement, à ouvrir et fermer les dispositifs de régulation du débit de fluide comme requis de sorte que le contenu d'un récipient soit transféré dans un autre récipient, le contenu du récipient étant isolé de l'environnement extérieur. Ce procédé est particulièrement utile pour la mise en culture des macrophages en atmosphère entièrement régulée, dans des conditions d'isolement hermétique à l'atmosphère extérieure.
Crotty Daphna
Danon David
Marks & Clerk
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