Mottet V
Laboratoire de Biologie, Institut Paoli-Calmettes (IPC) Marseille.
Chirurgie. 1994;120(2):100-3.
Growth cartilage were cultured in order to determine whether cartilage growth could replace bone loss and generate autologous bone capable of further growth. An abundant quantity of mature functional chondrocytes is required after culture. Chondrocytes were isolated from a cartilage fragment by enzyme digestion. Trypsin, then collagenase were used as a function of the number of chondrocytes obtained per gram of growth cartilage. The optimal culture conditions were determined on the basis of three parameters: the density of cell seeding and the type and composition of the culture medium. The kinetics of chondrocyte growth and their fixation on different support media as well as their functional capacity was determined in different media according to the following conditions: seeding at 200,000 or 300,000 chondrocytes/cm2 on type 1 collagen and in Hamm-F12 medium without supplementation with growth factors. The first trials of cryopreservation of the chondrocytes after culture gave promising results. Yields, expressed as the percentage of viable cells varied from 91 to 98% depending on the cryoprotective solution used.
培养生长软骨以确定软骨生长是否能够替代骨质流失并生成能够进一步生长的自体骨。培养后需要大量成熟的功能性软骨细胞。通过酶消化从软骨碎片中分离软骨细胞。根据每克生长软骨获得的软骨细胞数量,依次使用胰蛋白酶和胶原酶。基于三个参数确定最佳培养条件:细胞接种密度以及培养基的类型和组成。根据以下条件在不同培养基中测定软骨细胞生长动力学及其在不同支持培养基上的固定情况以及它们的功能能力:在1型胶原上以200,000或300,000个软骨细胞/cm²接种,并在不添加生长因子的Hamm-F12培养基中培养。培养后软骨细胞冷冻保存的首次试验取得了有前景的结果。根据所使用的冷冻保护溶液不同,活细胞百分比表示的产量在91%至98%之间变化。