METHOD FOR ISOLATION OF CARDIOMYOCYTES FROM HUMAN HEART TISSUE

摘要

FIELD: medicine.;SUBSTANCE: invention relates to the field of medicine, it can be used in biophysical and biological laboratories. The method includes sampling the biopsy, placing it in a buffer solution containing no Ca²⁺, grinding the biopsy, followed by transferring to a pure buffer solution of the same composition, mixing, then transferring the fragments of the biopsy to an enzymatic buffer containing a mixture of enzymes, one of which is collagenase, incubation at 37°C with stirring, followed by removal of the supernatant and repeated incubation in a fresh solution of the enzymatic buffer in the same mode. Then incubation is carried out in an enzymatic buffer containing only collagenase, followed by filtration of the cell suspension, centrifugation, then, removal of the supernatant and the introduction of a pure buffer solution containing no Ca²⁺, re-suspension of the precipitated cardiomyocytes, and the introduction of an aqueous solution of CaCl₂. The biopsy is ground to a size of 1-2 mm, and as an enzymatic buffer from a mixture of enzymes; a mixture of type 4 collagenase at a concentration of 1.02-1.42 mg/ml and pronase at a concentration of 0.17-0.2 mg/ml is used. An enzymatic buffer containing only type 4 collagenase is used with the same concentration of 1.02-1.42 mg/ml, while each incubation is carried out for 7-13 minutes. Incubation in an enzymatic buffer containing only collagenase is carried out at least 3 times for 7-15 minutes, and centrifugation is carried out at 800 rpm for 1-3 minutes. ;EFFECT: proposed method is simpler, more economical and provides for production of isolated viable cardiomyocytes from a human heart biopsy.;1 cl, 3 ex