人纤维环细胞与骨髓间充质干细胞Transwell共培养研究

目的探讨人的纤维环（AF）细胞与骨髓间充质干细胞（BMSCs）在Transwell共培养后,对AF细胞的细胞增殖和细胞外基质合成的影响,以及BMSCs是否具有向AF细胞分化的能力。方法分别对5例年龄在40岁以下脊柱手术患者的纤维环组织和骨髓标本进行分离、培养AF细胞和BMSCS,分别吸取第三代的AF细胞和BMSCs,按1 1的细胞比例分别植于Transwell共培养系统中,下室植入BMSCs,上室植入AF细胞。并且建立3个细胞培养组,即BMSCs与AF细胞共培养组（实验组）,BMSCs单独培养组和AF细胞单独培养组作为对照组。培养3周后,运用倒置相差显微镜观察细胞的形态变化;分别用HE染色、Ⅰ型胶原免疫组化染色和流式细胞仪对两种细胞鉴定;运用荧光定量PCR法检测各细胞培养组蛋白聚糖Aggrecan、Ⅰ型胶原蛋白mRNA及Sox9的表达变化。结果两种细胞在分离培养后经过鉴定确定为AF细胞和BMSCs。在倒置相差显微镜观察下,共培养组中AF细胞和BMSCs在细胞增殖数量和速度上均高于各自的单独培养组;共培养组中AF细胞和BMSCs的蛋白聚糖Aggrecan、Ⅰ型胶原蛋白mRNA及Sox9的表达均较各自的单独培养组高（P〈0.05）。结论通过BMSCs与AF细胞Transwell共培养,BMSCs可以促进AF细胞增值和细胞外基质的合成;同时通过共培养,BMSCs具有向AF细胞分化的可能。

The experimental study of human annulus fibrosus cell and bone marrow mesenchymal stem cells co-culture by transwell

Objective Discusses the human annulus fibrosus(AF) cell and the bone marrow mesenchymal stem cells(BMSCs) co-culture in Transwell on AF cells cell proliferation and extracellular matrix expression,and the BMSCs whether it has the ability to AF cell differentiation.Method For respectively between five the following ages of 40 years old spinal surgery patient AF cells and BMSCs separation,training.The third passage of BMSCs and AF cell were harvested and placed into Transwell co-culture system at a ratio of 1...