Poly-HEMA对大鼠剑突软骨细胞体外培养的影响  

The isolation and culture of rats xiphoid chondrocytes

作　　者：颉玉欣[1] 李亚[1] 郭晓华[1] 李国慧[1] 赵维丽[1] 张素领[1] 

机构地区：[1]河北医科大学第三医院实验中心,石家庄市050051

出　　处：《河北医药》2009年第18期2369-2371,共3页Hebei Medical Journal

摘　　要：目的研究多聚2-羟乙基甲基丙烯酸脂(Poly-HEMA)对体外培养大鼠剑突软骨细胞的生物学特性的影响,对细胞进行纯化过程,并作进一步的细胞形态学及细胞特性鉴定。方法无菌条件下取Wistern大鼠剑突,采用剪碎-酶消化法分离单个软骨细胞,台盼蓝染色鉴定细胞活性。将分离得到的细胞分别接种在预先用Poly-HEMA包被的平皿和普通的平皿内,并将普通平皿的细胞进行差速贴壁。结果从剑突软骨可获得大量软骨细胞,每克软骨可获得软骨细胞3.5×107个细胞,活性率为95.23%。软骨细胞在Poly-HEMA包被的平皿内培养使细胞始终呈悬浮状态生长,并能长期保持软骨细胞的特性。在普通平皿内经2次差速贴壁得到的软骨细胞纯度较高,但不转接Poly-HEMA包被的平皿内,细胞经过几次传代,即失去软骨细胞原有的特性。结论从剑突可获得大量软骨细胞,使用Poly-HEMA试剂体外培养的软骨细胞,其软骨细胞能长期以悬浮状态生长,并能使软骨细胞长期保持原有的生物学特性,为体外实验及组织工程学的研究提供更优质种子细胞,为临床的进一步研究提供理论基础。Objective To investigate the effect of poly-2-hydroxyethylmethacrylate（poly-HEMA） on the biological characteristics of rats xiphoid chondrocytes during the isolation and culture in vitro, to isolate the cells and identify the cells＇ morphous and characteristics. Methods The xiphoids of Wistern rats were taken out under sterile condition and to cut them, then to isolate single chondroeyte by enzyme digestion, and trypan blue staining was used to identify cells＇ viability. The isolated chondrocytes were implanted in culture dishes with poly-HEMA and without poly-HEMA. The cells in dishes without poly-HEMA were attached in different time points. Results A great of chondrocytes could be acquired from Xiphoid, the number of chondrocytes gained from wet xiphoid cartilage was （3.1 ± 0.5 ） × 10^7/g with a viability of 95.23% Chondrocytes in culture dishes coated with poly HEMA could grow in the suspension. The poly-HEMA could keep chondrocytes primary morphology and biological character. Chondrocytes could be purified after differential attachment twice in common dishes. But the chondrocytes lost primary characteristics after several passages. Conclusion A great of chondrocytes can be got from xiphoid, and the chondrocytes can grow in suspension and keep primary characteristics for a long time when they are cultured with poly-HEMA in vitro. In this way, good seed cells are provided for experiment in vitro, and for the study of tissue engineering, and for clinical study.

关 键 词：细胞培养 软骨细胞 Poly—HEMA 组织工程 

分 类 号：R-332[医药卫生]