不同方法体外培养软骨细胞的活力、功能及成软骨能力

The cartilage-formed potential of chondrocytes after cultured by various methods

目的探讨不同体外培养方法所培养软骨细胞活力、表型及成软骨能力的差异。方法取股骨髁关节面软骨,经不完全酶消化法消化后,与未完全消化的软骨块置入预涂多聚赖氨酸的培养瓶中培养;以传统法对照,用倒置显微镜、免疫组化等方法进行观测;并将多次传代后的软骨细胞悬液注入裸鼠腋部皮下,观察其成软骨能力。结果改良法培养的细胞活性率可达100%;经多次传代的软骨细胞仍保持初代软骨细胞的表型特征,并可观察到软骨的特有的细胞-胶原纤维几何构象;移植到裸鼠皮下后可形成软骨样组织。结论改良法是一种较方便、有效的培养法。

Objective To improve a simple and convenient method of human articular chondrocyte culture.Investigate it's chondrogenic potential after 7 passager in vitro.Methods The chondrocyte was obtained by incomplete enzyme digestion of cartilage from amputated human joint. It was seeded with small piece of cartilage in 25ml culture flasks which has coated with polylysine previously.After 7 passage, It was tested by Toluidine blue dine the glycoamine.Immunohistochemical method the collagen.Ⅱ and the growth by phase contrast microscopy. Results The percent of viable chondrocytes cultured by modified method with trypan blue staining was 100%.Chondrocytes still keep chondrocyte's phenotype after 7 passeger.The characteristic geometry of cell collagen fibrous can be seen by microscopy.After subcutaneous injected into armpit of naked rat, it can formed cartilage like tissue. Conclusions The modified method is a simple and convenient method