Human Stem Cell Group, Diabetes Transplant Unit, Prince of Wales Hospital, University of New South Wales, Sydney, Australia.
J Biomed Mater Res A. 2010 Feb;92(2):683-92. doi: 10.1002/jbm.a.32401.
Human embryonic stem cells (hESCs) have previously been cultured on three dimensional (3D) biodegradable polymer scaffolds. Although complex structures were formed from the hESCs, very little is known about the mechanism of adhesion of these cells to the surfaces of the scaffolds. In this study, we achieved the efficient adhesion of pluripotent hESCs to 3D poly(lactic-co-glycolic acid) (PLGA) scaffolds based on our data from a novel two dimensional (2D) model that imitates the surface properties of the scaffolds. In the 2D model, single cell preparations of pluripotent hESCs adhered efficiently and predominantly to PLGA surfaces coated with laminin in comparison to collagen I, collagen IV, or fibronectin-coated surfaces. Flow cytometry analysis revealed that almost all of the pluripotent single cells expressed the integrin alpha 6, with a small percentage also expressing alpha 3ss1, which facilitates adhesion to laminin. This data was then translated into the 3D environment, with the efficient binding of single pluripotent hESCs to PLGA scaffolds coated with laminin. The utility of this system was shown by the directed differentiation of single hESCs seeded within laminin-coated scaffolds toward the endoderm lineage.
人胚胎干细胞（hESCs）先前已在三维（3D）可生物降解聚合物支架上培养。尽管从 hESCs 形成了复杂的结构，但对于这些细胞与支架表面的粘附机制知之甚少。在这项研究中，我们根据模仿支架表面特性的新型二维（2D）模型的数据，实现了多能 hESCs 对 3D 聚（乳酸-共-乙醇酸）（PLGA）支架的有效粘附。在 2D 模型中，与胶原蛋白 I、胶原蛋白 IV 或纤维连接蛋白涂层表面相比，多能 hESCs 的单细胞制剂有效地且主要粘附到涂有层粘连蛋白的 PLGA 表面。流式细胞术分析显示，几乎所有的多能单细胞都表达整合素 alpha 6，一小部分也表达 alpha 3ss1，这有利于与层粘连蛋白的粘附。然后将该数据转化为 3D 环境，单细胞多能 hESCs 有效地结合到涂有层粘连蛋白的 PLGA 支架上。通过在涂有层粘连蛋白的支架内播种的单个 hESCs 向内胚层谱系的定向分化证明了该系统的实用性。
