Xie You-zhuan, Zhu Zhen-an, Tang Ting-ting, Dai Ke-rong, Lu Jian-xi, Pierre Hardouin
Department of Orthopaedic Surgery, Ninth People's Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200011, China.
Zhonghua Yi Xue Za Zhi. 2006 Jun 20;86(23):1633-7.
To investigate the feasibility of using perfusion culture bioreactor for bone mesenchymal stem cell proliferation in large scale beta-tricalcium phosphate (beta-TCP) scaffold.
In the dynamic perfusion culture group, the porous beta-TCP cylindrical scaffolds combined with the sheep mesenchymal stem cells were continuously perfused with the complete alpha-MEM medium by a peristaltic pump for 1, 2 and 4 weeks. While in the static culture group, the hybrid constructs were immersed in the medium without perfusion for 2 and 4 weeks. The cell proliferation and distribution were examined by the daily glucose consumption, the cell viability and undecalcified histological study.
The daily glucose consumption increased with time. The increase was much more evident in the first 2 weeks than in the last 2 weeks. The daily glucose consumption was higher in the dynamic culture group than in the static culture group. The cell viability also increased with time. It was higher in the dynamic culture group. In comparison to 2-week culture, the cell viability was significantly higher after 4-week culture in the dynamic culture group (P < 0.05), while it was not significantly different after 4-week culture in the static culture group (P > 0.05). Under dynamic perfusion culture, the mesenchymal stem cells survived and proliferated through the scaffolds. However, the mesenchymal stem cells survived and proliferated only in the peripheral pores of the scaffolds under static culture. Histomorphometrical study indicated that there were much more cells in dynamic culture group than in the static group. The cell/pore rate was not significantly different between 2-week static culture and 4-week static culture (P > 0.05). However, the cell/pore rate was significantly higher after 4-week dynamic culture than after 2-week dynamic culture (P < 0.05).
Perfusion culture permitted the persistent nutrition supply and gas exchange into the centre of large scaffold. This perfusion bioreactor makes the mesenchymal stem cells survive and proliferate through a large three-dimensional scaffold.
探讨使用灌注培养生物反应器在大尺寸β-磷酸三钙（β-TCP）支架中促进骨间充质干细胞增殖的可行性。
在动态灌注培养组中，将多孔β-TCP圆柱形支架与绵羊间充质干细胞相结合，通过蠕动泵用完全α-MEM培养基连续灌注1、2和4周。而在静态培养组中，将混合构建体浸入培养基中，不进行灌注，培养2和4周。通过每日葡萄糖消耗、细胞活力和非脱钙组织学研究来检测细胞增殖和分布情况。
每日葡萄糖消耗随时间增加。在前2周的增加比后2周更明显。动态培养组的每日葡萄糖消耗高于静态培养组。细胞活力也随时间增加。动态培养组的细胞活力更高。与2周培养相比，动态培养组在4周培养后细胞活力显著更高（P<0.05），而静态培养组在4周培养后无显著差异（P>0.05）。在动态灌注培养下，间充质干细胞在支架中存活并增殖。然而，在静态培养下，间充质干细胞仅在支架的周边孔隙中存活并增殖。组织形态计量学研究表明，动态培养组的细胞比静态组多得多。2周静态培养和4周静态培养之间的细胞/孔隙率无显著差异（P>0.05）。然而，4周动态培养后的细胞/孔隙率显著高于2周动态培养后（P<0.05）。
灌注培养能够持续向大尺寸支架中心提供营养供应和气体交换。这种灌注生物反应器使间充质干细胞能够在大型三维支架中存活并增殖。
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