Jouve J L, Cottalorda J, Mottet V, Frayssinet P, Petit P, Bollini G
Department of Pediatric Surgery, Hôpital Timone Enfants, Marseille, France.
J Pediatr Orthop B. 1998 Apr;7(2):174-8. doi: 10.1097/01202412-199804000-00017.
An original and reliable technique to culture growth plate chondrocytes was developed to obtain an abundant amount of mature and functional chondrocytes. Growth plates were provided from the epiphysis of 3-week-old rabbits. Isolation of the chondrocytes was optimized by the use of trypsin and collagenase. The culture was realized according to the following conditions: seeding at 20,000 or 30,000/cm2 on type I collagen substrate and in Ham F-12 medium without a supplementation of glucose or growth factors. After 7 days of culture, the implantation was to be carried out. Different implantation substrates were evaluated in vivo. Agar turned out to be the only substrate to provide strong and healthy chondrocytes 21 days after the grafting. Then implantation was tested on large iliac resections in rabbits to check whether an enchondral ossification occurred with the culture. Poor results were obtained because of an early disappearance of the cultured chondrocytes. In an other experimentation, the culture was implanted into surgically created defects in the growth plate area. In this case, the culture did produce an epiphysiodesis. However, the 6-week postoperative histological examination showed that the implant remained viable, continued to maintain a proteoglycanrich matrix, and began to organize in ordered columns of mature chondrocytes.
为了获得大量成熟且功能正常的软骨细胞,开发了一种原始且可靠的培养生长板软骨细胞的技术。生长板取自3周龄兔子的骨骺。通过使用胰蛋白酶和胶原酶优化软骨细胞的分离。培养按照以下条件进行:在I型胶原底物上以20000或30000个/cm²的密度接种,置于不含葡萄糖或生长因子补充剂的Ham F-12培养基中。培养7天后进行植入。在体内评估了不同的植入底物。结果发现,琼脂是唯一在移植21天后能提供强壮且健康软骨细胞的底物。然后在兔子的大髂骨切除术中进行植入测试,以检查培养物是否会发生软骨内成骨。由于培养的软骨细胞过早消失,结果不佳。在另一项实验中,将培养物植入生长板区域手术造成的缺损处。在这种情况下,培养物确实导致了骺板固定。然而,术后6周的组织学检查显示,植入物仍然存活,继续维持富含蛋白聚糖的基质,并开始组织成有序的成熟软骨细胞柱。