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目的探讨旋转式生物反应器是否能促进骨髓来源的间充质干细胞(MSCs)的快速扩增。方法将MSCs细胞放入盛有MyelocultTM培养液生物反应器中动态培养,并在MyelocultTM培养液中补充一些因子,如干细胞因子(SCF),白介素3和6(IL-3,IL-6)。检测和比较经反应器处理前后的MSCs的表型,增生和分化能力的变化。结果在生物反应器中培养8d,总细胞数、Stro-1+CD44+CD34-间充质干细胞和CD34+CD44+Stro-1-造血干细胞分别增加了9、29和8倍。实验研究结果显示,生物反应器扩增的间充质干细胞表达了原始间充质干细胞的标记物内皮因子(SH2)和波形蛋白,却没有发现与细胞系的分化有关的标记物,包括骨钙素(成骨指标),II型胶原(成软骨指标)和C/EBPα(成脂指标)。生物反应器组的细胞集落生成效率(成纤维细胞集落/天)是对照组(无生物反应器组)的1.44倍。经诱导后,生物反应器扩增的间充质干细胞可以分化成成骨细胞,软骨细胞和脂肪细胞,其分化能力与未经生物反应器处理的MSCs无区别。结论旋转式生物反应器结合改良的MyelocultTM培养液可用于快速扩增间充质干细胞。
Objective To investigate whether a rotating bioreactor can promote the rapid expansion of bone marrow-derived mesenchymal stem cells (MSCs). Methods MSCs were cultured in a bioreactor containing Myelocult medium and supplemented with some factors such as stem cell factor (SCF), interleukin 3 and 6 (IL-3, IL-6) in MyelocultTM medium. The phenotypes, proliferation and differentiation of MSCs before and after the reactor treatment were detected and compared. Results After cultured for 8 days in the bioreactor, the total number of cells, Stro-1 + CD44 + CD34-MSCs and CD34 + CD44 + Stro-1- hematopoietic stem cells increased 9, 29 and 8 times respectively. Experimental results show that bioreactor-derived mesenchymal stem cells express markers of primitive mesenchymal stem cells, endoglin (SH2) and vimentin, but no markers associated with cell lineage differentiation, including bone calcium (Osteogenetic index), type II collagen (cartilage index), and C / EBPα (adipogenic index). The colony forming efficiency (fibroblast colonies / day) of the bioreactor group was 1.44 times that of the control group (no bioreactor group). After induction, the bioreactor-expanded mesenchymal stem cells can differentiate into osteoblasts, chondrocytes and adipocytes, and their differentiation ability is not different from that of MSCs without bioreactor treatment. Conclusion Rotary bioreactor with improved MyelocultTM broth can be used to rapidly expand mesenchymal stem cells.