Isolation and Cultivation of Human Articular Chondrocytes

我們對15例因膝關節疾病或外傷切除的關軟骨進行了人關節軟骨細胞的分離與培養。應用酵素消化法分離細胞，用加有血清的F12培養液進行培養。本文並研究了血清與鹼性纖維細胞生長因子(basic fibroblast growth factor , bFGF)對培養的軟骨細胞生長的作用。血清濃度為1-30%時刺激軟骨細胞之生長。bFGF明顯刺激軟骨細胞之生長，當濃度為1-100ng/ml時其作用呈劑量-效應模式。軟骨細胞在加有10%胎牛血清的F12培養液中生長良好。在2-6個月培養期內可傳代多次，平均細胞數增加2702倍，總分裂次數達11次。用加有bFGF(10ng/ml)的培養液時，總分裂倍數可增加至24次。免疫化學研究(應用抗S-100抗體)顯示所有培養均為純軟骨細胞培養。本項研究顯示可用本文敘述之方法建立成年人關節骨細胞培養。培養的軟骨細胞可用於在體外研究軟骨細胞之生理與病理，並可用於自體軟骨細胞移植以治療關節軟骨缺損。

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並列摘要

Adult human articular chondrocytes were isolated from surgically excised articular cartilage from 15 patients suffering from trauma or disease of the knee. CelJs were isolated with an enzymatic digestion method and cultured with F12 medium supplemented with serum. The effects of serum and basic fibroblast growth factor (bFGF) on the growth of cultured chondrocytes were studied. Serum stimulated the growth of chondrocytes at concentrations from 1 - 30%. bFGF significantly stimulated the growth of chondrocytes in a dose-dependent manner at concentrations from 1 - 100 ng/ml. Chondrocytes grew well in F12 medium supplemented with 10% fetal bovine serum (FBS). These cultured chondrocytes could be passaged for many generations and revealed an average of 2, 702-fold increase of cell numbers during 2 - 6 months period (cumulative population doublings = 11 times). Cumulative population doublings increased to 24 times in cell cultured with medium supplemented with bFGF (10 nglml). Immunocytochemical study using anti-S-l 00 antibodies demonstrated that these cultures were pure cell cultures of chondrocytes. We have demonstrated that cell cultures of adult human articular chondrocytes can be established with these methods. Cultured chondrocytes provide 3n in vitro model system for studying the physiology and pathology of human articular chondrocytes and may be used for autologous transplantation of chondrocytes to treat articular cartilage defects.