METHOD FOR ISOLATION OF CHONDROCYTES Russian patent published in 2019 - IPC G01N33/48 C12N5/77 

FIELD: medicine.

SUBSTANCE: invention relates to medicine, in particular to a method for isolating chondrocytes. Method for isolating chondrocytes involves collecting material, cutting cartilage from the bone, cutting the cartilage into fragments, then the fragments of cartilage obtained are placed in a filtering device, incubated in trypsin solution, the resulting suspension of chondrocytes is centrifuged, the supernatant is discarded, and the resulting precipitate of chondrocytes is washed with a solution of phosphate buffer PBS 2 times, the first fraction of chondrocytes is sampled, then the unlysed cartilage fragments in the filtering device are incubated in a collagenase type II solution, the resulting precipitate of chondrocytes is washed with a solution of phosphate buffer PBS 2 times, the second fraction of chondrocytes is sampled, then the first and second chondrocyte fractions are combined, then the non-lysed cartilage fragments are removed from the filtering device and incubated in a DMEM/F12 culture medium containing FBS, penicillin, streptomycin, amphotericin B, then the above-described culture medium is discarded and a solution of type II collagenase, obtained after precipitation of chondrocytes by centrifugation in the second stage, is added, incubated, the resulting suspension of chondrocytes is centrifuged, the resulting supernatant containing type II collagenase is discarded, and the obtained precipitate of chondrocytes is washed with a solution of phosphate buffer PBS 2 times, the third fraction of chondrocytes is selected and combined with the first and second fractions of chondrocytes, then the resulting suspension of chondrocytes is plated on culture plastic for adhesion cultures in DMEM/F12 containing FBS, penicillin, streptomycin, amphotericin B, obtaining the primary cell population of the zero passage.

EFFECT: claimed method allows to increase the yield of chondrocytes.

1 cl, 1 ex