三维载体培养下离心力对骨髓间充质干细胞增殖的影响

Effects of centrifugal force on proliferation of bone marrow mesenchymal stem cells in the three-dimensional carrier

目的:观察在不同大小离心力作用下三维载体上骨髓间充质干细胞(BMSCs)的生长情况。方法:取大鼠第3代BMSCs种植于胶原海绵支架,分为对照组(0r/min)、200r/min组、500r/min以及800r/min组;实验组给予相应转速的离心力刺激,对照组同步置于除离心外相同的环境中;利用CM-Dil活细胞膜染色观察BMSCs与海绵支架的相容性,检测不同时相下端粒酶浓度,H-E染色评价BMSCs的生长情况。结果:BMSCs种植后存活,沿胶原海绵支架表面和空洞生长;端粒酶浓度在第15天内均递增,在第57天内均下降,且200r/min和500r/min组增高值>对照组,800r/min组<对照组;H-E染色显示200r/min和500r/min组细胞数量比对照组明显增多,以500r/min组最多,而800r/min组明显减少。结论:较低转速(200r/min与500r/min)的离心力可促进BMSCs增殖,而过高转速(800r/min)的离心力抑制BMSCs增殖,且在500r/min干预5d时细胞的生长活性最强。

Objective： To observe the growth of bone mesenchymal stem cells （BMSCs） cultured in the three-dimensional carrier under different centrifugal forces. Methods： The third-generation BMSCs were seeded into the collagen sponges and divided into control group, 200 r/min group, 500 r/min group and 800 r/min group with each experimental group being given corresponding centrifugal force. The control group was exposed to the same environment except for the centrifugal force. The compatibility of BMSCs with collagen sponge was observed by CM-Dil staining and the growth of BMSCs was assessed by telomerase concentration detection at different time points （1 d, 3 d, 5 d, 7 d） and H-E staining of paraffin section on the 7~ day. Results： BMSCs survived in the collagen sponges and proliferated well. Telomerase concentration of all groups gradually increased from day I to day 5 and decreased from day 5 to day 7. The concentration of 200 r/min group and 500 r/min group was higher than the control group, while that of the 800 r/min group was lower than the control group. H-E staining showed that the cell number of 200 r/min group and 500 r/min group was significantly higher than the control group, with the highest in the 500 r/rain group. However, the cell number of 800 r/rain group was significantly lower than the control group. Conclusion： Lower rotation speed （200 r/min and 500 r/min） promotes proliferation of BMSCs, while higher rotation speed （800 r/rain） inhibits the proliferation. The growth activity of BMSCs is the strongest in the 500 r/min group on the 5th day.