Three‐dimensional culture of mouse bone marrow cells within a porous polymer scaffold: effects of oxygen concentration and stromal layer on expansion of haematopoietic progenitor cells
Three‐dimensional culture of mouse bone marrow cells within a porous polymer scaffold: effects of oxygen concentration and stromal layer on expansion of haematopoietic progenitor cells
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DOI：
10.1002/term.295
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发表时间：
2011-02
影响因子：
3.3
通讯作者：
Hirotoshi Miyoshi;M. Murao;N. Ohshima;T. Tun
中科院分区：
文献类型：
--
作者：
Hirotoshi Miyoshi;M. Murao;N. Ohshima;T. Tun
为了建立造血祖细胞（HPC）和红系细胞的体外扩增方法，采用多孔聚乙烯醇缩甲醛（PVF）树脂作为支架进行小鼠骨髓细胞的三维（3D）培养。在这些培养物中，研究了氧浓度和与基质细胞共培养对HPC和红系细胞扩增的影响。当骨髓细胞在3D条件下培养时，HPC和红系细胞在不补充外源性细胞因子的情况下扩增，与基质细胞的存在无关。相反，在作为对照的单层培养物中观察到HPC或红系细胞的轻微扩增，表明使用PVF支架的3D培养物在扩增HPC和红系细胞方面比单层培养物好得多。在缺氧条件下，与不含基质细胞的培养物相比，骨髓基质细胞允许红系细胞和HPC以更高的细胞密度进行3D培养，并且启动3D共培养后HPC和红系细胞扩增的持续时间延长。在低氧条件下，这些细胞的数量在整个培养期间增加至3周，尽管在常氧条件下2周后数量减少。总之，证实了在缺氧条件下造血细胞与基质细胞的3D共培养方法在扩增HPC和红系细胞中是有效的，并且该方法似乎可用于开发造血细胞的离体扩增方法。版权所有© 2010约翰威利父子有限公司.
To establish an ex vivo expansion method of haematopoietic progenitor cells (HPCs) and erythroid cells, three‐dimensional (3D) cultures of mouse bone marrow cells were performed, employing a porous polyvinyl formal (PVF) resin as a scaffold. In these cultures, the effects of oxygen concentration and co‐cultures with stromal cells on the expansion of HPCs and erythroid cells were investigated. When bone marrow cells were cultured under 3D conditions, HPCs and erythroid cells expanded without supplementation of exogenous cytokines, irrespective of the presence of stromal cells. On the contrary, slight expansion of HPCs or erythroid cells was observed in monolayer cultures as controls, indicating that the 3D cultures using the PVF scaffold were far better in expanding HPCs and erythroid cells than the monolayer cultures. Under hypoxic conditions, bone marrow stromal cells allowed for a 3D culture of erythroid cells and HPCs at higher cell densities compared to cultures without stromal cells, and the duration of the expansion of HPCs and erythroid cells after initiating the 3D co‐cultures was prolonged. The number of these cells increased throughout the culture period up to 3 weeks under hypoxic conditions, although the number decreased after 2 weeks under normoxic conditions. In conclusion, the 3D co‐culture method of haematopoietic cells with stromal cells under hypoxic conditions was confirmed to be effective in expanding HPCs and erythroid cells, and this method seemed to be useful for developing an ex vivo expansion method for haematopoietic cells. Copyright © 2010 John Wiley & Sons, Ltd.