旋转细胞培养系统培养的山羊软骨细胞

 目的 探讨用旋转细胞培养系统（RCCS）进行大动物软骨细胞扩增的可行性。方法 将单层培养的具有正常表型的中国山羊软骨细胞和Cytodex-3微载体放入RCCS的培养容器中，加入DMEM 培养基进行培养。在细胞培养的第3、6、9、12 天,于倒置显微镜下观察Cytodex-3微载体表面软骨细胞生长形态及增殖变化情况，并对收获的软骨细胞进行蕃红花"O"染色和Ⅰ、Ⅱ型胶原免疫组织化学染色分析。结果 软骨细胞贴附于微载体表面生长，细胞初期呈球形、半球形凸起，逐渐向周围伸展，随时间的延长，贴附于微载体的细胞逐渐增多。收获的软骨细胞蕃红花"O"染色呈强阳性，Ⅰ型胶原染色呈阴性，Ⅱ型胶原染色则呈强阳性。结论 利用RCCS可简便快速地在体外扩增羊软骨细胞，培养的软骨细胞具有正常的表型，能特异性分泌软骨组织特有的基质成分。

Goat chondrocytes produced in vitro by rotary cell culture system

Objective To investigate the feasibility of amplifying big animal chondrocytes in vitro by using rotary cell culture system (RCCS). Methods Monolayer cultured goat articular chondrocytes, which had normal phenotype, were co-cultured with cytodex-3 microcarriers in RCCS added with DMEM. After 3, 6, 9, and 12 days, the growth and proliferation of the chondrocytes on the surface of the cytodex-3 microcarriers were observed under a phase-contrast microscope. The chondrocytes were collected, and stained with safranin O, and type Ⅰ or Ⅱ collagen antibodies for histological and immunocytochemical analysis. Results The articular chondrocytes grew rapidly covering the surface of cytodex-3 microcarriers. The cells clustered together forming round or hemispheroid protrusion, and then expanded around the microcarriers. The number of chondrocytes on the surface of cytodex-3 microcarriers increased with time. The harvested chondrocytes were stained positive for safranin O, negative for typeⅠcollagen, and strong positive for type Ⅱ collagen. Conclusions RCCS is a quick and convenient method for amplifying goat chondrocytes. The amplified chondrocytes, which had normal phenotypes, can secrete the specific stromal components of cartilage.