应用微重力旋转生物反应器培养小鼠脂肪干细胞的初步实验研究

Proliferation and Differentiation of Mouse Adipose Stem Cells in Rotary Cell Culture System

目的:观察微重力旋转培养系统(Rotary Cell Culture System,RCCS),对小鼠脂肪干细胞增殖的影响,以寻求一种更有效的促进干细胞扩增的方法。方法:从小鼠的脂肪组织中提取分离、培养脂肪干细胞(ADSCs),并对脂肪干细胞进行流式鉴定后,利用活细胞观察法、Dil免疫荧光标记法、扫描电镜法观察微重力旋转三维培养系统对脂肪干细胞增殖的影响;通过与平面二维培养作对比,血小板计数法记录细胞的增殖情况,并绘制生长曲线。结果:两组的细胞倍增时间具有统计学意义(P<0.05),模拟微重力旋转三维培养系统较传统平面二维培养系统,脂肪干细胞增殖更明显,生长速度更快。结论:模拟微重力旋转三维培养系统更有利于脂肪干细胞的增殖生长,为后期利用脂肪干细胞修复受损涎腺提供一种更快捷有效的扩增方法。

Objective： To find a more effective method to promote amplification of the stem cells, through observing the influence of the 3D microgravity rotation system （RCCS） to the development of adipose stem cell proliferation. Methods： Extracted and separated the fat tissue from the mice, cultivating adipose stem cells （ADSCs）, and using flow cytometer to identify the fat cells, using living cells observation, Dil immunofluorescence mark method, scanning electron microscopy （SEM） method to observe the influence of 3D microgravity rotating culture system to the fat stem cell proliferation; use platelet method to record the cell proliferation and draw the growth curve. Results： We found that cell doubling time had statistical significance contrast to the two-dimensional culture method （P〈 0.05）. Compared to the traditional two-dimensional culture system, the 3D microgravity rotating culture system is more apparent in cell proliferation and cells are growing faster. Conclusion： 3D simulated microgravity rotating culture system is more advantageous to fat cells proliferation growth, which is a more efficient amplification method for tissue engineering.