用组织培养法保存软骨活性

Study on vital cartilage preservation using tissue culture method

目的 :观察用组织培养法保存软骨块活性的可行性 .方法 :取成年新西兰白兔全厚层喉甲状软骨 ,切成大小约5mm× 2mm× 1mm小块后用RPMI16 4 0培养液进行组织培养 ,并与 4 0 g·L-1甲醛和 9mL·L-1生理盐水所保存的软骨在保存 2 4h ,5 ,10 ,2 0和 30d时分别用倒置显微镜、HE和AB/PAS切片染色及台盼蓝排斥试验进行软骨活性比较 .结果 :用 4 0mL·L-1甲醛和生理盐水所保存的软骨分别于 2 4h及10d后基本失去活性 ,软骨细胞及基质变性坏死 ,而用RPMI16 4 0培养液所保存软骨在培养 30d后仍保持较好活性 ,软骨基质强度无明显变化 ,软骨细胞活性保持在 75 %以上 .结论 :与甲醛等化学保存方法相比 ,用组织培养法能较长期地保存软骨块活性 。

AIM: To study the possibility of using tissue culture for vital cartilage preservation. METHODS: The thyroid cartilage of 9 New Zealand white rabbits were totally dissected. The cartilage samples were cut into 5 mm×2 mm×1 mm fragments. The fragments (total 18) were randomly divided into three groups, and then they were separately preserved in the medium RPMI1640, 40 g·L -1 formaldehyde and 9 mL·L -1 saline solution. The viabilities of the cartilage grafts were compared after 24 h, 5, 10, 20 and 30 d, using microscope and trypan blue dye exclusion test. RESULTS: Cartilage kept in the saline solution and formaldehyde showed a quick decrease of viability. The cartilage lost its viability after 24 h kept in formaldehyde and 10 d in the saline solution. In contrast, cartilage preserved in the RPMI1640 medium retained its viability during the whole 30 day storage time. CONCLUSION: Compared with chemical preservation procedures such as formaldehyde, the tissue culture method can successfully keep the viability of cartilage grafts for 30 d. It may offer a new vital cartilage preservation method for clinical treatment.