兔骨髓间充质干细胞在Cytodex 3微载体悬浮培养系统中的贴附条件优化

Optimization of Attachment Conditions for Rabbit Mesenchymal Stem Cells in Cytodex 3 Microcarrier Culture Systems

骨髓间充质干细胞（Mesenchymal stem cells，MSCs）在组织工程及基因治疗中具有重要的应用价值，为实现MSCs体外动态扩增，本研究采用微载体培养技术，考察兔MSCs接种入Cytodex3微载体悬浮培养系统后的贴附情况。在低速连续搅拌条件下，MSCs接种后8～12h贴附达到平衡，贴附率仅为16．7％±1．1％；采用间歇搅拌方式、50％培养体积接种、降低贴附期培养基中血清浓度均使MSCs贴附率有显著提高；在四种培养基体系中．MSCs在aMEM中的贴附率比DMEM中平均高39．8％。与常规条件相对照，采用优化后的接种方法．贴附率从26．6％提高到65．5％，细胞总体扩增倍数从2．01提高到4．50。本研究结果为实现MSCs在微载体悬浮培养系统中大量扩增奠定了基础。

Mesenchymal stem ceils are of great value in tissue engineering and genetic engineering. To study the in vitro dynamic expansion of mesenchymal stem ceils by microcarrier technology, we began the research with the investigation of attachment of rabbit mesenchymal stem ceils on cytodex 3 microcarriers after inoculation. The result showed a poor attachment efficiency of 16.7%±1.1% under general conditions, so the attachment efficiency must be increased through the optimization of inoculation conditions. Intermittent stirring, inoculation in ,50% of the final culture volume and reduction of the fetal bovine serum concentration at inoculation all led to notable increases in attachment efficiency. Ratio of rMSCs attached in aMEM cultures was 39.8 % higher than that of DMEM＇s on the average. When all these optimal conditions were adopted, the attachment efficiency （65.5%） was significantly higher than that （26. 6%） under general conditions; accordingly the final growth extents of rMSCs were 4. SO and 2. 01, respectively. Therefore, the optimization of attachment conditions promoted the expansion of rMSCs on microcarriers.