Stimulated cell standards

G - Physics – 01 – N

Patent

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G01N 33/96 (2006.01)

Patent

CA 2730191

Methods for producing stimulated, positive and negative control reference standard for monitoring intracellular cytokine levels and cytokine release in test samples by stimulating cells to produce cytokines in the presence of a cytokine release inhibitor, fixing the stimulated cells with a fixative such as paraformaldehyde, washing to remove excess fixatives and freeze-drying the stimulated, fixed cells. Methods for producing labelled reference standards for cell proliferation assays are also disclosed, in which proliferation-competent mammalian cells, isolated from a human or animal body are labelled with a label, such as a dye, that is divided between daughter cells during cell proliferation (e.g. carboxyfluorescein succinimidyl ester), the cells are stimulated to proliferate, the proliferated cells are fixed by addition of a fixative and then preserved by freeze drying or cryopreservation.

Linvention concerne des procédés de préparation de standards de référence stimulés, de contrôles positifs et négatifs, pour surveiller les niveaux intracellulaires de cytokine et la libération de cytokine dans des échantillons tests par stimulation de cellules pour produire des cytokines en présence dun inhibiteur de la libération de cytokines, fixation des cellules stimulées avec un fixateur tel que le paraformaldéhyde, lavage pour éliminer lexcès de fixateur et lyophilisation des cellules stimulées fixées. Linvention concerne également des procédés de préparation de standards de référence marqués pour des essais de prolifération cellulaire, selon lesquels des cellules mammaliennes compétentes en matière de prolifération, isolées dun corps humain ou animal, sont marquées avec un marqueur, tel quun colorant, qui est divisé entre les cellules filles pendant la prolifération cellulaire (p. ex. ester succinimidylique de carboxyfluorescéine). Les cellules sont stimulées pour proliférer, les cellules proliférées sont fixées par ajout dun fixateur, puis conservées par lyophilisation ou cryoconservation.

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