猪耳廓软骨细胞外分泌基质的检测

Glycosaminoglycan secretion in porcine auricle chondrocytes culture

目的 通过体外传代培养猪耳廓软骨细胞 ,并连续检测软骨细胞形成外分泌基质的能力 ,寻找出最适宜接种的种子细胞。方法 通过Ⅱ型胶原酶消化法获得高活性的软骨细胞 ,定量接种 6× 10 6个细胞于培养皿 ,每周传代 1次 ,连续 5周 ,留取所有培养液用阿利新蓝法测定软骨细胞外分泌基质糖胺多糖 (GAG) ;利用光镜观察原代及传代软骨细胞的生长情况。结果 第 1,2 ,3 ,4,5周软骨细胞外分泌基质GAG平均含量分别为 :612 ,12 3 5 ,14 62 ,113 8和 710 μg。 结论 体外培养的第 2代软骨细胞功能和活性最佳 ,传代第 2周是最佳接种时间。

Objective To determine the optimal time of cultured chondrocytes for innoculation by quantitating the secretion of glycosaminoglycan(GAG). Methods\ Chondrocytes were cultured in plate(6×10 6 cells) and passed on each week, for 5 consecutive weeks. Secretion of GAG was determined by DMS- GAG colorimetry assay from week 1 to weekr 5. Results\ The average GAG was 612, 1 235 , 1 462 ,1 138 and 710 μg respectively from week 1 to week 5.Conclusion\ GAG synthesized by cultured chondrocytes increases significantly at week 3, and decreases after week 4. Cultured chondrocytes were most active during week 2, optimal of innoculation.