METHOD OF REMOVING CELLS FROM CULTURE SURFACE DURING SUBCULTURE OF MULTIPOTENT MESENCHYMAL STROMAL CELLS Russian patent published in 2010 - IPC C12N5/735 C12N9/48 

FIELD: chemistry; biochemistry.

SUBSTANCE: invention relates to cellular biology, particularly to a method of removing multipotent mesenchymal stromal cells from a culture surface during subculture, and can be used when culturing stem cells with their subsequent use in cellular therapy. After removing the culture medium, a first portion of a 0.075% solution of accutase is added to the culture of multipotent mesenchymal stromal cells in amount of 5 ml per 25 cm² of the culture surface. The enzyme is aspirated after a few seconds. A second portion of a 0.075% solution of accutase is added right after removing the first portion of enzymes. Incubation is carried out for 2-3 seconds, after which the 0.075% solution of accutase is aspired again. Subsequent incubation is carried out in accutase enzyme vapour for 2 minutes at 37°C.

EFFECT: change in conditions of enzymatic treatment of the culture cells during culturing lies in that, removal of cells in accutase enzyme vapour increases output of living cells.

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