三维培养骨软骨复合体融合研究

目的 探讨三维培养体系中构建骨软骨复合体的可行性.方法 取10周以内流产胚胎组织原代培养人胚胎干细胞,按1×1O7个/ml细胞数植入自制的微流控支架三维培养系统,分别用相应细胞因子诱导分化成骨细胞和软骨细胞.体外培养3 d后植入裸鼠体内,8周后取出标本进行组织学观察.结果 支架骨端孔隙率为45%,孔径为125～150 μm;软骨端孔隙率为85%,孔径150～250 μm.组织学检查显示再造组织形成了骨和软骨复合组织,但是两者之间仍有界限,完全融合失败.结论 三维培养骨软骨复合体,可成功再造出骨软骨复合组织,但是完全融合尚需增加支架软骨端的孔隙率和孔径,以及延长体内培养的时间.

Abstract：

Objective To explore the feasibility of construction of bone cartilage complex in the three-dimensional culture. Methods The abortive embryos within 10 weeks were obtained for primary culture of human embryonic stem cells (hESCs). The hESCs about 1 × 107/ml were implanted into microfluidic three-dimensional culture system. hESCs were induced to differentiate into osteoblasts and chondrocytes by cytokine respectively. After culture in vitrofor 3 days, the system was transplanted in nude mouse.Eight weeks after implantation, the specimens were harvested and examined histologically. Results In the microfluidic three-dimensional culture system, the porosity of bony part was 45%, and the aperture was 125-150 μm; the porosity of chondral part was 85%, and the aperture was 150-250 μm. In the newly formed tissue, it demonstrated the formation of bone cartilage complex tissue under the microscopy. However, there was still a boundary between the two tissues. The perfect fusion of bone and cartilage became failure. Conclusion In the experiment, bone cartilage complex tissue can be reconstructed in three-dimensional culture system, but the perfect fusion needs to increase the porosity and the aperture of chondral part, together with culture time elongation in vivo.

Fusion of bone cartilage complex in the three-dimensional culture

Objective To explore the feasibility of construction of bone cartilage complex in the three-dimensional culture. Methods The abortive embryos within 10 weeks were obtained for primary culture of human embryonic stem cells (hESCs). The hESCs about 1 × 107/ml were implanted into microfluidic three-dimensional culture system. hESCs were induced to differentiate into osteoblasts and chondrocytes by cytokine respectively. After culture in vitrofor 3 days, the system was transplanted in nude mouse.Eight weeks after implantation, the specimens were harvested and examined histologically. Results In the microfluidic three-dimensional culture system, the porosity of bony part was 45%, and the aperture was 125-150 μm; the porosity of chondral part was 85%, and the aperture was 150-250 μm. In the newly formed tissue, it demonstrated the formation of bone cartilage complex tissue under the microscopy. However, there was still a boundary between the two tissues. The perfect fusion of bone and cartilage became failure. Conclusion In the experiment, bone cartilage complex tissue can be reconstructed in three-dimensional culture system, but the perfect fusion needs to increase the porosity and the aperture of chondral part, together with culture time elongation in vivo.