Determination of optimal transport conditions for biopsies of human articular cartilage as well as for suspensions of cultured chondrocytes used in autologous transplantation

Autologous transplantation of chondrocytes is currently being promoted as a novel approach for the treatment of deep cartilage lesions. Briefly, the m ethod involves enzyme-mediated release of chondrocytes from cartilage biops ies, the expansion of cells by in vitro cultivation and their re-implantati on into the defect. The success of this technique depends on many factors i ncluding transport conditions for both, cartilage biopsies from the operati ng hall to the laboratory and the return transport of final suspension of c ultured chondrocytes. To determine the extent of cellular damage in biopsie s, chondrocytes were enzymatically isolated following a few days of tissue preservation in different tissue culture media. The proportion of dead cell s was assessed by Trypan blue staining and counting. The viability was not dependant of the type of the medium used and remained approximately 50% in all samples, even after 72 h. To develop optimal conditions for transport of final chondrocyte suspension , isolated cells were firstly grown in monolayer cultures. Cell suspensions in media with different additives were injected into special glass contain ers used for the transport and left at 4oC or 25oC for up to 3 days. During this period every 24 h the samples were taken and viability as well as apo ptosis levels were assessed. Viability of cells in suspensions at 25 degree sC decreased significantly and became inadequate already after 48 h. In con trast to that, the proportion of viable cells at 4 degreesC remained above 80% even after 48 h. In the majority of the samples, culture medium contain ing serum and vitamin C provided the best conditions for long-term preserva tion of chondrocytes.