玻璃化冻存骨髓基质干细胞实验探究.doc

玻璃化冻存骨髓基质干细胞实验探究[摘要]目的：骨髓基质干细胞（BMSCs）是构建组织工程化骨主要的种子细胞，实现BMSCs的深低温保存对骨组织工程具有重要意义。目前，玻璃化冻存是最有发展前景的冻存方法，因此希望通过改进玻璃化液和预处理条件来提高BMSCs的玻璃化冻存效果。方法：采用不同组成及浓度的玻璃化液在不同的预处理条件下对第2（P2）代成骨诱导的犬骨髓基质干细胞（cBMSCs）进行玻璃化冻存实验，通过复苏后的细胞存活率来选择一种理想的玻璃化液及适合的预处理条件，并在此基础上再与常规冻存的实验结果进行比较，同时考虑冻存过程对细胞成骨活性的影响。结果：将VS226作为玻璃化液，在0℃/5min的预处理条件下玻璃化冻存P2代成骨诱导的cBMSCs，与常规冻存后的细胞存活率相比无统计学差异，且冻存过程对此细胞的成骨能力没有影响。结论：采用VS226来玻璃化冻存BMSCs，可实现为骨组织工程提供大量种子细胞的目的。 [关键词]深低温保存；玻璃化冻存；骨组织工程；骨髓基质干细胞 [中图分类号]Q813.1 [文献标识码]A [文章编号]1008-6455（2009）03-0318-05 Experimental study of vitrification preservation for BMSCs YIN Hong-yu1, SUN Jian3, LIU Guang-peng3,CUI Lei2,3,CAO Yi-lin1,2,3 (1. Research Center Of Plastic Surgery Hospital, Chinese Academy of Medical Sciences Peking Union Medical College, Beijing 100041, China; 2.Department of Plastic and Reconstructive Surgery, Shanghai Ninth People’s Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200011, China; 3.Shanghai Tissue Engineering Research and Development Center, Shanghai 200234, China.) Abstract: Objective Bone marrow stromal cells (BMSCs) have become the main cell source for tissue-engineered bone. There is of great significance for bone tissue engineering to cryopreserved BMSCs. At present, vitrification preservation is the most promising methods of cryopreservation for cells. The effect of vitrification method for BMSCs was wished to enhance by improving the vitrified solution and the pretreatment condition. Methods Using the different composition and concentration of vitrification fluid, the freezing experiments on osteogenically induced canine bone marrow stromal cells (cBMSCs) at passage 2 (P2) were performed under the different preconditioning. According to the cellular survival rate after rewarming, the ideal vitrification media and optimal pretreatments were chosen to cryopreserve the cells in order to compare with the experimental result of slow-freezing. Simultaneously, the influence of vitrification protocol on osteogenic activit