TUMOR-CELL HAPTOTAXIS ON COVALENTLY IMMOBILIZED LINEAR AND EXPONENTIAL GRADIENTS OF A CELL-ADHESION PEPTIDE
TUMOR-CELL HAPTOTAXIS ON COVALENTLY IMMOBILIZED LINEAR AND EXPONENTIAL GRADIENTS OF A CELL-ADHESION PEPTIDE
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DOI：
10.1016/0012-1606(89)90159-0
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发表时间：
1989-09-01
影响因子：
2.7
通讯作者：
SCHNAAR, RL
中科院分区：
文献类型：
--
作者：
BRANDLEY, BK;SCHNAAR, RL
细胞沿粘附基质梯度向上移动已被认为是在发育和转移过程中指导细胞迁移的机制。对这一假设（趋触性）的批判性评价得益于使用生物学相关粘附分子的可量化、稳定的基质梯度。我们报告的聚丙烯酰胺表面的共价衍生化与定量梯度的九肽含有粘合剂的精氨酸-甘氨酸-天冬氨酸序列。通过用B16 F10鼠黑素瘤细胞均匀接种衍生化表面来研究细胞迁移。在8小时内，细胞的梯度重新分布显着，较高的细胞密度被发现在凝胶位置具有较高的固定化肽密度。相反，接种在具有均匀浓度的粘附肽的对照凝胶上的细胞没有重新分布。在无血清和含血清培养基中，梯度上发生再分布。均匀密度肽衍生凝胶的实验表明，在梯度上的再分布不是由于优先初始细胞附着或优先生长在较高密度的固定化肽，但必须是由于细胞易位。固定化肽的指数梯度上的细胞迁移到凝胶表面上对应于最高固定化肽密度的位置，而相同肽的线性梯度上的细胞迁移到中间肽密度的位置。这些数据表明，B16 F10细胞响应固定化肽密度的比例变化，而不是绝对的变化，这意味着一个传感机制，利用适应。这些结果表明，（1）小粘附肽的梯度足以产生细胞群的再分布，以及（2）可以产生受控的可定量基质梯度，并用于探测该行为的潜在细胞机制。
The movement of cells up an adhesive substratum gradient has been proposed as a mechanism for directing cell migration during development and metastasis. Critical evaluation of this hypothesis (haptotaxis) benefits from the use of quantifiable, stable substratum gradients of biologically relevant adhesion molecules. We report covalent derivatization of polyacrylamide surfaces with quantifiable gradients of a nonapeptide containing the adhesive Arg-Gly-Asp sequence. Cell migration was studied by seeding derivatized surfaces evenly with B16F10 murine melanoma cells. Within 8 hr, cells on gradients redistributed markedly; higher cell densities were found at gel positions having higher immobilized peptide densities. In contrast, cells seeded on control gels with uniform concentrations of adhesive peptide did not redistribute. Redistribution occurred on gradients in both serum-free and serum-containing media. Experiments with uniform density peptide-derivatized gels demonstrated that redistribution on gradients was not due to preferential initial cell attachment or preferential growth on the higher density of immobilized peptide, but must have been due to cell translocation. Cells on exponential gradients of immobilized peptide migrated to a position on the gel surface corresponding to the highest immobilized peptide density, while cells on linear gradients of the same peptide migrated to a position of intermediate peptide density. These data suggest that the B16F10 cells respond to proportional changes in immobilized peptide density rather than to absolute changes, implying a sensing mechanism which utilizes adaptation. These results demonstrate that (1) a gradient of a small adhesive peptide is sufficient to generate redistribution of cell populations and (2) controlled quantifiable substratum gradients can be produced and used to probe the underlying cellular mechanisms of this behavior.