Designer self-assembling peptide nanofiber scaffolds for adult mouse neural stem cell 3-dimensional cultures.-猫眼课题宝

DOI：

10.1371/journal.pone.0000119

发表时间：

2006-12-27

期刊：

PloS one

影响因子：

3.7

通讯作者：

中科院分区：

综合性期刊3区

文献类型：

作者：

Gelain F;Bottai D;Vescovi A;Zhang S

来源链接：

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1762423/?report=classic

生物医学研究人员越来越意识到传统的二维组织细胞培养系统的局限性，包括涂膜培养皿、多孔板和载玻片，无法充分解决细胞生物学、癌症生物学和神经生物学中的许多关键问题，如三维微环境、三维梯度扩散、三维细胞迁移和三维细胞-细胞接触相互作用。为了充分了解细胞在三维人体中的行为，重要的是开发一个控制良好的三维细胞培养系统，其中每一种成分都是已知的。在这里，我们报告了使用设计肽纳米纤维支架和小鼠成体神经干细胞的三维细胞培养系统的发展。我们将几个功能基序，包括细胞粘附、分化和骨髓归巢基序，连接到一个自组装肽RADA16 （Ac-RADARADARADARADA-COHN2）上。这些功能化肽经过自组装形成类似于Matrigel的纳米纤维结构。在细胞培养过程中，细胞完全嵌入支架的三维环境中。两种含有骨髓归巢基序的肽支架在常规细胞培养基中不需要额外的可溶性生长因子和神经营养因子，显著提高了神经细胞的存活率。在这些设计支架中，具有β-Tubulin+、GFAP+和Nestin+标记的细胞群与在Matrigel上培养的细胞群相似。基因表达谱阵列实验显示，选择性基因表达可能参与了神经干细胞的粘附和分化。由于合成的肽本质上是纯净的，并且许多所需的功能细胞基序很容易结合，这些设计肽纳米纤维支架为多种组织细胞提供了一个有前途的可控三维培养系统，并且在一般分子和细胞生物学中也很有用。

Biomedical researchers have become increasingly aware of the limitations of conventional 2-dimensional tissue cell culture systems, including coated Petri dishes, multi-well plates and slides, to fully address many critical issues in cell biology, cancer biology and neurobiology, such as the 3-D microenvironment, 3-D gradient diffusion, 3-D cell migration and 3-D cell-cell contact interactions. In order to fully understand how cells behave in the 3-D body, it is important to develop a well-controlled 3-D cell culture system where every single ingredient is known. Here we report the development of a 3-D cell culture system using a designer peptide nanofiber scaffold with mouse adult neural stem cells. We attached several functional motifs, including cell adhesion, differentiation and bone marrow homing motifs, to a self-assembling peptide RADA16 (Ac-RADARADARADARADA-COHN2). These functionalized peptides undergo self-assembly into a nanofiber structure similar to Matrigel. During cell culture, the cells were fully embedded in the 3-D environment of the scaffold. Two of the peptide scaffolds containing bone marrow homing motifs significantly enhanced the neural cell survival without extra soluble growth and neurotrophic factors to the routine cell culture media. In these designer scaffolds, the cell populations with β-Tubulin+, GFAP+ and Nestin+ markers are similar to those found in cell populations cultured on Matrigel. The gene expression profiling array experiments showed selective gene expression, possibly involved in neural stem cell adhesion and differentiation. Because the synthetic peptides are intrinsically pure and a number of desired function cellular motifs are easy to incorporate, these designer peptide nanofiber scaffolds provide a promising controlled 3-D culture system for diverse tissue cells, and are useful as well for general molecular and cell biology.