Bioreactor expansion of human adult bone marrow-derived mesenchymal stem cells
Bioreactor expansion of human adult bone marrow-derived mesenchymal stem cells
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DOI：
10.1634/stemcells.2005-0591
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发表时间：
2006-09-01
期刊：
影响因子：
5.2
通讯作者：
Li, Gang
中科院分区：
文献类型：
--
作者：
Chen, Xi;Xu, Haibo;Li, Gang
在造血干细胞（HSC）移植过程中补充间充质干细胞（MSC）可减少并发症，如移植物抗宿主病，导致造血快速恢复。为了满足这种临床需求，需要一种快速的MSC扩增方法。在本研究中，我们研究了使用旋转生物反应器系统从分离的骨髓单个核细胞扩增MSC的可行性。在旋转生物反应器中用含有补充因子组合的Myelocult培养基培养细胞，所述补充因子包括干细胞因子和白细胞介素-3和白细胞介素-6。培养8天后，总细胞数、Stro-1（+）CD 44（+）CD 34（-）MSC和CD 34（+）CD 44（+）Stro-1（-）HSC分别增加了9倍、29倍和8倍。经生物反应器处理的细胞的集落形成效率-成纤维细胞/天是未经生物反应器处理的细胞的1.44倍。生物反应器扩增的MSC显示原始MSC标志物内皮糖蛋白（SR 2）和波形蛋白的表达，而与谱系分化相关的标志物，包括骨钙蛋白（骨生成），H型胶原（软骨生成）和C/EBP-α（CCAAT/增强子结合蛋白-α）（脂肪生成），未检测到。在诱导后，生物反应器扩增的MSC能够分化成成骨细胞、软骨细胞和脂肪细胞。我们的结论是，旋转生物反应器与本研究中报道的改良Myelocult培养基可用于快速扩增MSC。
Supplementation of mesenchymal stem cells (MSCs) during hematopoietic stem cell (HSC) transplantation alleviates complications such as graft-versus-host disease, leading to a speedy recovery of hematopoiesis. To meet this clinical demand, a fast MSC expansion method is required. In the present study, we examined the feasibility of using a rotary bioreactor system to expand MSCs from isolated bone marrow mononuclear cells. The cells were cultured in a rotary bioreactor with Myelocult medium containing a combination of supplementary factors, including stem cell factor and interleukin-3 and -6. After 8 days of culture, total cell numbers, Stro-1(+)CD44(+)CD34(-) MSCs, and CD34(+)CD44(+)Stro-1(-) HSCs were increased 9-, 29-, and 8-fold, respectively. Colony-forming efficiency-fibroblast per day of the bioreactor-treated cells was 1.44-fold higher than that of the cells without bioreactor treatment. The bioreactor-expanded MSCs showed expression of primitive MSC markers endoglin (SR2) and vimentin, whereas markers associated with lineage differentiation, including osteocalcin (osteogenesis), type H collagen (chondrogenesis), and C/EBP-alpha (CCAAT/enhancer-binding protein-alpha) (adipogenesis), were not detected. Upon induction, the bioreactor-expanded MSCs were able to differentiate into osteoblasts, chondrocytes, and adipocytes. We conclude that the rotary bioreactor with the modified Myelocult medium reported in this study may be used to rapidly expand MSCs.