快速扩增羊软骨细胞的生物学特性观察

To observe the goat's chondrocytes biological characteristics amplified by a quick way in vitro

目的：观察在旋转生物反应器内微载体技术快速扩增的羊软骨细胞生物学特性。方法：将培养的羊软骨细胞应用Cytodex-3微载体在旋转生物反应器（RCSS）内，进行动态培养，应用倒置显微镜和扫描电子显微镜对微载体表面的软骨细胞进行动态观察，对收获的软骨细胞进：行细胞计数、蕃红花“0”染色、Ⅱ型胶原的细胞免疫组化分析。结果：关节软骨细胞于1d内贴附于Cytodex-3微载体表面，到12d，细胞密度可达最初接种的15—17倍，经蕃红花“0”染色为阳性、Ⅱ型胶原染色则呈强阳性。结论：利用微载体细胞培养技术可简便快速地在体外扩增羊软骨细胞。

Objective： To observe the goats chondrocytes biological characteristics amplified quickly in large scale by RCSS using microcarriers technique. Methods：Articular chondrocytos were harvested from goat by sequential digestion with trypsin and collagenase, and then grown in RCSS bioreactor culture system, which contained the Cytodex-3 microcarriers in the culture medium （DMEM）. Count the number of chondrocytes at different time. Growth of chondrocytes on Cytodex-3 microcarriers was observed dynamically under phase contrast microscope. Immunocytochemical analysis was performed for type Ⅱ collagen. Results： The articular chondrocytes attached to the surface of Cytodex-3 microcarriers in the first day. The density of chondrocytes increased 15-17 times in later stage of culture as compared with the initial density. The harvested chondrocytos had stained intensively for collagen type Ⅱ. Conclusions：By micro-carrier and RCSS culture of chondrocytes can yield a large quantity of goat cells within a short time, which will be of benefit to banking active and cartilage cells for reconstruction of impaired cartilage by way of tissue engineering.