Isolation and culturing of chondrocytes from different sources

Abstract

The investigation was aimed to establish conditions for isolation and culturing cells of cartilaginous tissue procured from different sources.

Methods of chondrocytes obtaining and cultureing were compared. The chondrocytes were procured from human fetal cartilaginous tissue and the spinal column of patients with idiopathic scoliosis, as well as intervertebral disks of a new-born dog backbone.

The investigations of obtaining and culturing of chondrocytes from different sources showed that using collagenase solutions during 12-16 h at 37°С with continuous interfusion allowed to receive cells suspension with more than 90% viability. The maximal amount of viable chondrocytes was derived while segregating cells using 0.15% collagenase. Нuman fetal cartilaginous tissue and that of dog intervertebral disks is being dissociated in collagenase for 5-8 h, tissue from the spinal column of patients with idiopathic scoliosis for 18-20 h.

While culturing human fetal chondrocytes had the best proliferating capability. The culture of chondrocytes from the spinal column of idiopathic scoliosis patients had low proliferation index and a short culturing time as compared with fetal culture.

Keywords

chondrocytes / cell culture / culturing / collagenase / tripsin / hyaluronidase

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