Insulin Diabetes Experimental Research Group, Faculty of Medicine and Health Sciences, University of Nottingham, United Kingdom.
J Biotechnol. 2009 Dec;144(4):304-12. doi: 10.1016/j.jbiotec.2009.08.008. Epub 2009 Aug 15.
This work investigates an application of the alginate encapsulation technology to the differentiation of embryonic stem (ES) cells into insulin-producing cells. It shows that the ES cells can efficiently be encapsulated within the alginate beads, retaining a high level of cell viability. The alginate encapsulation achieves approximately 10-fold increase in the cell density in the culture, in comparison to the two-dimensional conditions, opening a potential benefit of the technology in large-scale cell culture applications. Manipulations of encapsulation conditions, particularly of the initial alginate concentration, allow the control over both the diffusion of molecules into the alginate matrix (e.g. differentiation factors) as well as control over the matrix porosity/flexibility to permit the proliferation and growth of encapsulated ES aggregates within the bead. Post-differentiation analysis confirms the presence of insulin-positive cells, as judged from immunostaining, insulin ELISA and RT-PCR analysis. The functionality of the encapsulated and differentiated cells was confirmed by their insulin production capability, whereby on glucose challenge the insulin production by the cells differentiated within alginate beads was found to be statistically significantly higher than for the cells from conventional two-dimensional differentiation system.
本工作研究了海藻酸钠包封技术在胚胎干细胞（ES 细胞）向胰岛素分泌细胞分化中的应用。结果表明，ES 细胞可以有效地被包封在海藻酸钠珠内，保持高细胞活力。与二维条件相比，海藻酸钠包封使细胞密度在培养中增加了约 10 倍，这为该技术在大规模细胞培养应用中带来了潜在的益处。包封条件的操作，特别是初始海藻酸钠浓度的操作，可以控制分子向海藻酸钠基质中的扩散（例如分化因子），并控制基质的孔隙率/柔韧性，从而允许包封的 ES 细胞聚集体在珠内增殖和生长。分化后分析通过免疫染色、胰岛素 ELISA 和 RT-PCR 分析证实了存在胰岛素阳性细胞。通过其胰岛素产生能力证实了包封和分化细胞的功能，其中在葡萄糖刺激下，发现海藻酸钠珠内分化的细胞的胰岛素产生在统计学上显著高于传统二维分化系统中的细胞。
