藻酸钠微球培养对兔髓核细胞生物学特性的影响

Culture of rabbit nucleus pulposus cells in alginate beads and study the effect of biological characteristics

目的探讨藻酸钠微球培养对兔椎间盘髓核细胞(NPCs)体外生物学特性的影响。方法4月龄健康新西兰大耳白兔6只,取髓核细胞原代培养,传代后分为实验组(藻酸钠微球培养)和对照组(平面培养),通过倒置相差显微镜对髓核细胞进行形态学观察,MTT法测定两组髓核细胞增殖情况,运用RT-PCR技术检测两组髓核细胞中Ⅱ型胶原和聚集蛋白聚糖的表达。结果经过2周的培养,两组髓核细胞增殖率无明显差异;藻酸钠微球培养组在Ⅱ型胶原和聚集蛋白聚糖的表达上均高于平面培养组(P<0.01或P<0.05)。结论藻酸钠微球培养法能促进髓核细胞中Ⅱ型胶原和聚集蛋白聚糖的表达,在维持其表型稳定方面优于平面培养法。

Objective To culture rabbit nucleus pulposus cells using alginate beads and plates,then detected the biological character in vitro.Methods We got the primary nucleus pulposus cells（NPCs）from 6 healthy New Zealand rabbits of 4-month-old,and then divided the cells into the experiment group（culture in alginate beads）and the control group（culture on plates）after passage.Cell morphology of NPCs was observed by inverted phase contrast microscope.MTT was used to determine the cells proliferation,and RT-PCR was applied to detect the expression of collagen Ⅱand aggrecan in NPCs.Results After two weeks cultivation,there was no significant difference in the cells proliferation rate of NPCs between the experiment group and the control group.But the expression of collagen Ⅱand aggrecan in the experiment group was superior to the control（P〈0.01 or P〈0.05）.Conclusion Culturing NPCs in alginate beads can promote the expression of collagen Ⅱand aggrecan,it was better than that on plates to maintain phenotypic stability of NPCs.