Stepwise Assembly of Micropatterned Co-cultures Using Photoresponsive Culture Surfaces and Its Application to Hepatic Tissue Arrays
Stepwise Assembly of Micropatterned Co-cultures Using Photoresponsive Culture Surfaces and Its Application to Hepatic Tissue Arrays
复制标题
DOI：
10.1002/bit.22253
复制
发表时间：
2009-06-15
影响因子：
3.8
通讯作者：
Kanamori, Toshiyuki
中科院分区：
文献类型：
--
作者：
Kikuchi, Kyoko;Sumaru, Kimio;Kanamori, Toshiyuki
细胞微图案化是一种将细胞放置在任意区域的方法，正在成为进行细胞生物学和组织工程的重要工具。传统的细胞图案化技术通常只允许在相同的培养表面上进行具有单一细胞类型的单一图案化。然而，今天的生物医学研究甚至需要复杂的制造方法，需要对多个细胞排列进行时空控制。在这里，我们介绍了原位细胞微图案化系统，该系统可以使用光响应性细胞培养表面(PRCS)实现逐步细胞图案化，其细胞粘附性可以通过紫外光照射而改变。为了展示其在组织工程中的应用，我们制作了一个模拟肝脏的组织阵列，并用实时荧光定量聚合酶链式反应对肝脏特异性基因的表达进行了定量。构建了由HepG2球体和Balb/3T3组成的图案化共培养体系，得到了细胞功能最好的最佳球体直径为150µm，经20天的图案化共培养，细胞色素P3A4的表达比常规培养的HepG2高50倍，比随机共培养的HepG2和Balb/3T3高20%。因此，PRCS和免掩模照射装置的结合显示了实验装置的多功能性，并被证明是生物医学研究的有力工具。生物技术。比昂斯。2009年；103：552-561。(C)2009年威利期刊公司。
Cell micropatterning, a method to place cells at arbitrary regions, is becoming an essential tool to conduct cell biology and tissue engineering. Conventional cell patterning techniques usually allow only single patterning with single cell type on the same culture surface. However, biomedical research today requires even sophisticated fabrication methods that require spatiotemporal control of multiple cell arrangements. Here we introduce in situ cell micropatterning system which enables stepwise cell patterning using a photoresponsive cell culture surface (PRCS) whose cell adhesiveness could be altered by the UV irradiation. To demonstrate an application to tissue engineering, a liver-mimic tissue array was fabricated and liver-specific gene expressions were quantified with real time PCR. Patterned co-culture systems composed of HepG2 spheroids with Balb/3T3 were fabricated, and the optimum spheroid diameter, which yielded the highest cellular functions, was determined to be 150 mu m. After 20 days of patterned co-culture of HepG2 spheroids and Balb/3T3, CYP3A4 expression increased 50-fold higher than conventionally cultured HepG2; CYP3A4 expression was 20% higher than randomly co-cultured HepG2 and Balb/3T3. Thus the combination of PRCS and the photomask-free irradiation apparatus showed the versatility of experimental setups and proved to be a powerful tool for biomedical studies. Biotechnol. Bioeng. 2009;103: 552-561. (C) 2009 Wiley Periodicals, Inc.