Micropatterned substrates: Approach to probing intercellular communication pathways
Micropatterned substrates: Approach to probing intercellular communication pathways
复制标题
DOI：
10.1021/ac0257400
复制
发表时间：
2002-09-15
影响因子：
7.4
通讯作者：
Porter, MD
中科院分区：
文献类型：
--
作者：
Takano, H;Sul, JY;Porter, MD
细胞间信号传导对于中枢神经系统（CNS）的正常发育和生理至关重要。为了研究这种信号，控制细胞在何时何地相互接触至关重要。确定用于细胞定位的过程是否对信号传导有影响也很重要。本文描述了一种技术，控制在体外细胞生长的位置，并证明该技术对细胞间信号传导的影响最小（如果有的话）。通过使用凝胶渗透法，制造聚（二甲基硅氧烷）模具作为模板的非粘附性琼脂（琼脂糖）上的细胞粘附膜（聚（L-赖氨酸））涂布的玻璃盖玻片微米级图案。该过程产生由明确的粘附性和非粘附性微区组成的表面。当将内皮细胞或星形胶质细胞铺在这些基质上时，内皮细胞或星形胶质细胞的汇合域在聚（L-赖氨酸）域上生长。星形胶质细胞和神经元的共培养也可以成功地用于在粘附结构域上形成交织的网络。此外，钙信号的研究表明，在这种模式下生长的星形胶质细胞保留了其天然的生理活性。这一结论是基于观察到的个别星形胶质细胞域内和相邻的，但空间上断开，星形胶质细胞域的钙波的传播速率。潜在的应用这些微图案化的基板作为平台，询问通信途径的中枢神经系统的关键组成部分进行了讨论。
Intercellular signaling is critical for the normal development and physiology of the central nervous system (CNS). To study such signaling, it is vital to control where and when the cells make contact with one another. It is also important to determine whether the process used for cell localization has an impact on signaling. This paper describes a technique that controls the location for cell growth in vitro and demonstrates that the technique has minimal (if any) impact on intercellular signaling. By using photolithographic methods, poly(dimethylsiloxane) molds were fabricated to function as templates for micrometer-level patterning of a nonadhesive agar (agarose) onto glass coverslips coated with a cell adhesive film (poly(L-lysine)). This process yields a surface composed of well-defined adhesive and nonadhesive microdomains. When endothelia or astrocytes are plated onto these substrates, confluent domains of endothelia or astrocytes grow on the poly(L-lysine) domains. Cocultures of astrocytes and neurons can also successfully be used to form interwoven networks on the adhesive domains. Moreover, studies of calcium signaling revealed that astrocytes grown on such patterns retain their native physiological activity. This conclusion is based on the observed propagation rate for calcium waves within individual astrocyte domains and across neighboring, but spatially disconnected, astrocyte domains. The potential to apply these micropatterned substrates as platforms for interrogating communication pathways in key components of the CNS is discussed.