Investigations of low-temperature storage of articular cartilage for transplantation

Isolated bovine articular cartilage chondrocytes and intact slices of cartilage were investigated to determine the effects of low-temperature cryopreservation on articular cartilage. Studies have focused on prefreezing conditions of cartilage, including the incubation medium and temperature of incubation, type and toxicity of the cryopreservative used, and the penetration of cryopreservative agents into cartilage cells. Cartilage freezing conditions were examined with respect to rate of freezing, controlled differential freezing rates, the ultimate storage temperature, and the time of storage. Cartilage thawing conditions were observed to ascertain the role of membrane osmotic stress during thawing and the effect of variable thawing rates on the viability of chondrocytes. Careful control of these variables can yield cartilage with cellular viability of over 50%. Optimum cryopreservation of viable cartilage should include prefreezing treatment with 7.5%-10% DMSO in nutrient medium, controlled slow freezing to -70 degrees, and rapid thawing in DMSO containing medium. A significant number of chondrocytes in deep-frozen cryopreserved articular cartilage can survive. The work recommends continued clinical use of deep-frozen cartilage.     

荧光原位杂交技术分析人结肠菌群方法研究

建立荧光原位杂交技术分析人体内结肠菌群的方法。取受试者新鲜粪便 ,选用 5种特异性的 16SrRNA寡核苷酸探针 ,检测粪便样本收集后的保存时间、温度 ,离心条件及样本固定液存放时间对杂交计数结果的影响。结果建立最佳实验条件为 :粪便样本收集后应尽快在 4℃下保存 ,放置时间不要超过 12小时即作处理 ;样本的适宜离心条件为 70 0g 2分钟 ;样本用多聚甲醛固定后在 - 80℃下存放时间不要超过 5个月。该方法具有较好的稳定性 ,可以有效地检出个体之间结肠菌群的差异。