人骨髓基质细胞软骨诱导分化及其与细胞载体体外复合  

The Human Bone Marrow Stromal Cells Differentiate into Chondroblast and Culture on PDLLA/Chitosan and Hydroxyapatite Scaffold in vitro

作　　者：胡军[1] 张爱斌[1] 陈永松[1] 杨佩璇[1] 宋建新[1] 刘晓岚 

机构地区：[1]汕头大学医学院第一附属医院骨科,广东汕头515041 [2]第一人民医院脊柱外科,湖南郴州423000

出　　处：《汕头大学医学院学报》2005年第3期135-138,F0002,共5页Journal of Shantou University Medical College

摘　　要：目的:观察出生后人骨髓基质细胞(hBMSCs)在体外培养条件下增殖与分化的特点,探讨其向成软骨方向分化及机制。研究诱导细胞体外与PDLLA/壳聚糖多孔材料复合。方法:密度梯度离心法进行hBMSCs体外培养。应用传五代细胞,经化学限定培养基诱导细胞,设实验对照组。采用倒置显微镜观察细胞增殖及形态变化,甲苯胺蓝染色观察诱导细胞合成细胞外基质中的蛋白多糖,RT-PCR检测Ⅱ型胶原mRNA的表达。将诱导的细胞分别与多孔羟基磷灰石、PDLLA/壳聚糖多孔材料复合,应用扫描电镜观察其在细胞载体表面的生长情况。结果:传五代hBMSCs经化学限定培养基诱导后转化成圆形肥大细胞,甲苯胺蓝染色阳性,Ⅱ型胶原mRNA表达与对照组比较有统计学意义。扫描电镜发现hBMSCs在PDLLA/壳聚糖多孔材料表面增殖良好并分泌大量细胞基质。结论:体外培养hBMSCs经化学限定培养基诱导后可向成软骨方向分化,可作为软骨组织工程种子细胞。PDLLA/壳聚糖多孔复合材料是组织工程良好的细胞载体,有利于细胞的黏附与增殖。Objective： To examine the property and proliferation ability of primary culture human bone marrow stromal cells（hBMSCs）, and to investigate the effect of hBMSCs differentiation into chondroblast. Experiment on the possibility of PDLLA/chitosan porous material as chondroblast matrix scaffold, step to reveal the role for PDLLA/ehitosan. Methods： hBMSCs were source of the bone marrow collected from healthy human donors and cultured according to Majumdar＇s method in vitro, the cells of fifth subculture were cultured by chemical definition medium, examined by invert microscope. Toluidine blue stain and RT-PCR were used to test the type Ⅱ collagen mRNA. To implant hBMSCs of fifth generation on PDLLA/chitosan and HA was observed by scanning electron microscope. Results： The hBMSCs could be differentiated into chondroblast, and RT-PCR showed they could synthesis type Ⅱ collagen. Scanning electron microscope showed the cells proliferated quickly in PDLLA/chitosan scaffold. Conclusion： hBMSCs can keep proliferation and differentiation ability in vitro, hBMSCs can differentiate into chondroblast by chemical definition medium, hBMSCs are a kind of ideal seed-cells for cartilage tissue engineering. The porous compound material of PDLLA/chitosan is an ideal scaffold in cartilage tissue engineering.

关 键 词：人骨髓基质细胞 成软骨分化 组织工程 细胞培养 

分 类 号：Q813.11[生物学—生物工程] R318.1[医药卫生—生物医学工程]