Department of Chemical Engineering, The Ohio State University, Columbus, 43210, USA.
Cytotechnology. 2003 Jan;41(1):23-35. doi: 10.1023/A:1024283521966.
Embryonic stem (ES) cells have indefinite self-renewal ability and pluripotency, and can provide a novel cell source for tissue engineering applications. In this study, a murine CCE ES cell line was used to derive hematopoietic cells in a 3-D fibrous matrix. The 3-D matrix was found to maintain the phenotypes of undifferentiated ES cells as indicated by alkaline phosphatase (ALP) activity and stage specific embryonic antigen-1 (SSEA-1) expression. In hematopoietic differentiation, cells from 3-D culture exhibited similar cell cycle distribution and SSEA-1 expression to those in the initial cell population. The Oct-4 expression was significantly down-regulated, which indicated the occurrence of differentiation, although the level was slightly higher than that in Petri dish culture. The expression of c-kit, cell surface marker for hematopoietic progenitor, was higher in the 3-D culture, suggesting a better-directed hematopoietic differentiation. Cells in the 3-D matrix tended to form large aggregates associated with fibers. For large-scale processes, a perfusion bioreactor can be used for both maintenance and differentiation cultures. As compared to the static culture, a higher growth rate and final cell density were resulted from the perfusion bioreactor due to better control of the reactor environment. At the same time, the differentiation capacity of ES cells was preserved in the perfusion culture. The ES cell culture in the fibrous matrix thus can be used as a 3-D model system to study effects of extracellular environment and associated physico-chemical parameters on ES cell maintenance and differentiation.
胚胎干细胞具有无限的自我更新能力和多能性，可以为组织工程应用提供新的细胞来源。在这项研究中，使用小鼠 CCE 胚胎干细胞系在 3D 纤维基质中衍生造血细胞。3D 基质被发现能够维持未分化 ES 细胞的表型，如碱性磷酸酶（ALP）活性和阶段特异性胚胎抗原-1（SSEA-1）表达。在造血分化中，来自 3D 培养的细胞表现出与初始细胞群体相似的细胞周期分布和 SSEA-1 表达。Oct-4 表达显著下调，表明发生了分化，尽管水平略高于 Petri 盘培养。造血祖细胞表面标志物 c-kit 的表达在 3D 培养中更高，表明造血分化的定向性更好。细胞在 3D 基质中倾向于与纤维形成大的聚集物。对于大规模过程，可以使用灌注生物反应器进行维持和分化培养。与静态培养相比，由于更好地控制反应器环境，灌注生物反应器可实现更高的生长速度和最终细胞密度。同时，ES 细胞的分化能力在灌注培养中得以保留。因此，纤维基质中的 ES 细胞培养可以用作 3D 模型系统，以研究细胞外环境和相关物理化学参数对 ES 细胞维持和分化的影响。
