In vitro matrix assembly induced by critical assembly concentration (CAC).
In vitro matrix assembly induced by critical assembly concentration (CAC).
复制标题
临界组装浓度 (CAC) 诱导的体外基质组装。
DOI：
10.1177/002215540205001114
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发表时间：
2002
期刊：
影响因子：
--
通讯作者：
McCarthy,KevinJ
中科院分区：
文献类型：
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作者：
Lauer,MarkE;McCarthy,KevinJ
L-2细胞是来源于卵黄囊壁层内胚层细胞的永生化细胞系，其负责产生Reichert膜，一种在大鼠妊娠期间产生的厚基底膜。虽然L-2细胞分泌基底层的所有主要成分，但它们在细胞培养中不组装稳健的基质。我们假设，L-2细胞在细胞培养中不能组装基质的原因是因为在标准细胞培养条件下，这种基质组装所需的基质组分浓度未达到临界缔合浓度（CAC）。为了限制分泌分子的扩散，同时保持营养丰富的环境使细胞茁壮成长，我们开发了一种技术，该技术使用透析膜来限制2孔板格式中的蛋白质扩散。这种技术允许L-2细胞组装一个强大的矩阵在短短24小时，继续形成至少72 hr. This技术可以解决一些细胞培养所施加的物理限制，并可以很容易地应用到其他细胞类型和培养基条件。
L-2 cells are an immortalized cell line derived from yolk sac parietal endoderm cells, which are responsible for the production of Reichert's membrane, a thick basement membrane produced during rat gestation. Although the L-2 cells secrete all the major components of the basal lamina, they do not assemble a robust matrix in cell culture. We hypothesized that the reason L-2 cells fail to assemble a matrix in cell culture is because the concentrations of matrix components necessary for this matrix assembly do not reach a critical association concentration (CAC) under standard cell culture conditions. To limit the diffusion of secreted molecules while maintaining a nutrient-rich environment for the cells to thrive, we developed a technique that uses a dialysis membrane to limit protein diffusion in a 2-well plate format. This technique permits L-2 cells to assemble a robust matrix in as little as 24 hr that continues to be formed for at least 72 hr. This technique may address some of the physical limitations imposed by cell culture and could be readily applied to other cell types and medium conditions.