软骨细胞在微载体中的培养和快速扩增

目的：探索在短期内获得大量成活率高、分化良好的兔关节软骨细胞的方法,方法：应用胰蛋白酶、胶酶消化的方法从新生新西兰兔关节软骨处分离、培养软骨细胞,并将获得的软骨细胞在旋转生物反应应（RCCS）内应用Cytodex-3微载体进行培养。应用倒置显微镜和扫描电镜对微载体表面的软骨细胞进行动态观察,并对收获的软骨细胞进行Ⅰ、Ⅱ型胶原的细胞免疫化学染色分析。结果：并节软骨细胞可快速贴附于Cytodex-3微载体表面,细胞伸展后生长加速,到培养后期,细胞密度可达最初接种的20倍。在微载体上收获的软骨细胞Ⅰ型胶原的免疫细胞化学染色呈阴性,Ⅱ型胶原染色则呈强阳性。结论：微载体细胞培养技术是一种简便、快速的体外细胞扩增方法,可为构体建组织工程化人工软骨提供大量软骨软件。

A quick way in vitro for amplification of chondrocytes

ObjectiveTo establish a quick way for acquiring welld if ferentiated chondrocytes in large scale. Methods: Articular chon drocytes were harvested from newborn NewZealand rabbits by sequential digestio n with trypsin and collagenase, and then grown in RCCS bioreactor culture syst em which contained the Cytodex3 microcarriers in the culture medium (DMEM).Growth of chondrocytes on Cytodex3 microcarriers was observed dynamically unde r phase contrast microscope and scanning electron microscope.Immunocytoc hemical analysis was performed for type~{"q~}collagen and type~{"r~}collagen. Re sultsThe articular chondrocytes attached rapidly t o the surface of Cytodex3 microcarriers.Quick growth of these cells was obs erved after they fully spread onto the microcarriers. The density of chondrocyte s increased 20 times in later stage of culture as compared with the initial dens ity. The harvested chondrocytes had no detectable staining for collagen type~{"q~}bu t stained intensively for collagen type~{"r~}. Conclusions Microcar rier culture of chondrocytes can yield a large quantity of cells within a short time,which will be of benefit to bank ing cartilage cells for reconstruction of impaired cartilage by way of tissue engineering.