永生化下颌骨髁突软骨细胞的微囊化研究

Microencapsulation of Immortalized Mandibular Condylar Chondrocytes

为了探讨微囊包裹软骨细胞在软骨组织工程中的适用性 ,根据气流切割原理采用海藻酸钠 -多聚赖氨酸 -海藻酸钠 ( APA)对永生化下颌骨髁突软骨细胞 ( Im mortalized mandibular condylar chondrocyte,IMCC)进行微囊包裹。用倒置显微镜观察、台盼蓝染色、细胞记数、HE染色、免疫组化等方法检测微囊的大小、细胞的生长及微囊内组织的软骨特性等情况。研究发现 ,IMCC可在微囊内存活 ,活细胞率 >80 % ,微囊直径平均 779μm。细胞数量随着培养时间的延长逐渐增多 ,约 2 0 d左右达到平台期 ,细胞在囊内呈簇样生长 ,高表达软骨特异的蛋白多糖和 型胶原。提示 IMCC可在微囊内形成类软骨组织样结构 。

To explore the possibility of microencapsulation of chondrocytes in cartilage tissue engineering,immortalized manibular condylar chondrocytes (IMCCs) were microencapsuled by Alginate-polylysine-alginate (APA) method, according to air pressure shearing model. Phase contrast microscopy, trypan blue staining exclusion, cell number counting, HE staining and immunohistochemistry method were used to observe the morphology of the microencapsules, the growth character of cells, cartilage characteristics, and so on. The results showed that IMCC could survive and grow in microencapsule, and the viability rate of cells is more than 80 per cent. The diameter of microcapsule is 779 μm in average. The number of cell increased with time, and cells went into platform in about 20 days. Cells grew in clusters and cartilage specific proteoglycans and type II collagen were highly expressed. It was concluded that IMCC could form cartilage-like tissue within microencapsulation, implying that microencapsule technique might be applicable to cartilage tissue engineering.