大鼠脂肪源性干细胞与三维打印明胶支架的相容性

Compatibility between adipose derived stem cells of rats and three dimensional printing gelatin scaffold

目的通过观察大鼠脂肪源性干细胞(ADSCs)与三维打印(3DP)及戊二醛交联的明胶支架的相容性,筛选出最适合细胞生长的支架孔径,为进一步构建组织工程化组织或器官提供实验依据。方法采用酶消化法分离提取大鼠ADSCs,并用流式细胞术和多向诱导分化的方法进行鉴定;然后与不同孔径的3DP明胶支架复合培养,并用扫描电子显微镜和透射电子显微镜观察其超微结构,细胞活力分析仪检测其存活率;用MTT法检测二维(2D)与三维(3D)培养对ADSCs细胞活力的影响。结果获得的ADSCs具有多向分化潜能,具备干细胞的基本特征。接种ADSCs到3DP明胶支架后,扫描电子显微镜可看到细胞呈椭圆形或纺锤形,区别于传统二维培养的梭形形态;透射电子显微镜可看到细胞在支架空隙内散在分布,细胞核、细胞器等结构清晰,表明其与支架的相容性良好;90μm孔径的支架细胞存活率最高;3D培养的方法更有利于维持ADSCs的活力。结论 ADSCs与3DP明胶支架的相容性良好,90μm孔径的支架最适合ADSCs的生长。

Objective To observie the compatibility of rat adipose-derived stem cells（ ADSCs） and three dimensional printing（ 3DP） gelatin scaffolds crosslinked with glutaraldehyde,the most suitable for cell growth aperture,in order to provide experimental basis for constructing tissue-engineered tissues or organs. Methods The enzyme digestion method was adopted to separate and extract rat ADSCs. Flow cytometry and multi-directional differentiation method were used for identification. The ADSCs were cultured with 3DP gelatin scaffolds of different aperturs. Scanning electron microscopy（ SEM） and transmission electron microscope（ TEM） were used to observe the ultra-microstructure. The cell vitality analyzer was used to detect its survival rate. MTT method was used to detect the effects of two dimensional（ 2D）and three dimensional（ 3D） cultural methods on ADSCs cell vitality. Results The seeding ADSCs had multi-directional differentiation potential,and had the basic characteristics of stem cells. After inoculation of ADSCs to 3DP gelatin scaffolds,cells were oval or spindle observed by SEM,which was different from the traditional 2D culture form. The cells scattered within the scaffold gap,the structures such as the nucleus and organelles were clear,which demonstrated a good compatibility with scaffold. The cell survival rate of 90μm aperture scaffold was the highest,and 3D cultural method was more advantageous to maintain the vitality of ADSCs. Conclusion The compatibility of ADSCs with 3DP gelatin scaffold is good,and 90μm aperture scaffold is most suitable for the growth of ADSCs.