Differentiation of human embryonic stem cells on three-dimensional polymer scaffolds
Differentiation of human embryonic stem cells on three-dimensional polymer scaffolds
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DOI：
10.1073/pnas.1735463100
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发表时间：
2003-10-28
影响因子：
11.1
通讯作者：
Langer, R
中科院分区：
文献类型：
--
作者：
Levenberg, S;Huang, NF;Langer, R
人类胚胎干细胞（hES）有望成为移植治疗的无限细胞来源。然而，控制它们的增殖和分化成复杂的、有活力的3D组织是具有挑战性的。在这里，我们研究使用生物可降解聚合物支架促进hES细胞的生长和分化，并形成三维结构。我们表明，复杂的结构与功能的各种提交的胚胎组织，在体外，通过使用早期分化的hES细胞，并进一步诱导其分化的支持性三维环境，如聚（乳酸-共-乙醇酸）/聚（L-乳酸）聚合物支架。我们发现hES细胞分化和组织化可以受到支架的影响，并受到生长因子如视黄酸、转化生长因子13、激活素A或胰岛素样生长因子的指导。这些生长因子分别诱导分化成具有发育神经组织、软骨或肝脏特征的3D结构。此外，观察到3D血管样网络的形成。当移植到严重的联合免疫缺陷小鼠，构建体继续表达特定的人类蛋白质在定义的分化结构，并出现招募和anastamose与宿主血管。这种方法提供了一种独特的培养系统，用于解决细胞和发育生物学中的问题，并提供了一种潜在的机制，用于创造可行的人体组织结构用于治疗应用。
Human embryonic stem (hES) cells hold promise as an unlimited source of cells for transplantation therapies. However, control of their proliferation and differentiation into complex, viable 3D tissues is challenging. Here we examine the use of biodegradable polymer scaffolds for promoting hES cell growth and differentiation and formation of 3D structures. We show that complex structures with features of various committed embryonic tissues can be generated, in vitro, by using early differentiating hES cells and further inducing their differentiation in a supportive 3D environment such as poly(lactic-co-glycolic acid)/poly(L-lactic acid) polymer scaffolds. We found that hES cell differentiation and organization can be influenced by the scaffold and directed by growth factors such as retinoic acid, transforming growth factor 13, activin-A, or insulin-like growth factor. These growth factors induced differentiation into 3D structures with characteristics of developing neural tissues, cartilage, or liver, respectively. In addition, formation of a 3D vessel-like network was observed. When transplanted into severe combined immunodeficient mice, the constructs continue to express specific human proteins in defined differentiated structures and appear to recruit and anastamose with the host vasculature. This approach provides a unique culture system for addressing questions in cell and developmental biology, and provides a potential mechanism for creating viable human tissue structures for therapeutic applications.