Baker S E, DiPasquale A P, Stock E L, Quaranta V, Fitchmun M, Jones J C
Department of Cell and Molecular Biology, Northwestern University Medical School, Chicago, Illinois 60611, USA.
Exp Cell Res. 1996 Nov 1;228(2):262-70. doi: 10.1006/excr.1996.0325.
The rat cell line 804G assembles an extracellular matrix which induces not only the rapid adhesion and spreading of epithelial cells but also the assembly of a cell-matrix attachment device called the hemidesmosome. The major component of this matrix is laminin-5. We have purified rat laminin-5 from medium conditioned by 804G cells. Epithelial cells which are co-incubated with medium supplemented with soluble laminin-5 adhere and spread rapidly. Furthermore, human carcinoma cells undergo a dramatic morphologic change in the presence of laminin-5 and form orderly arrays resembling epithelial sheets. Soluble rat laminin-5 is selectively incorporated into an insoluble matrix of epithelial cells in vitro, since rat-specific laminin-5 antibodies stain cell-substrate contacts. Addition of medium containing soluble laminin-5 to explanted, human corneal rims induces assembly of hemidesmosomes, important cell-matrix attachment devices. Furthermore, rat-specific laminin-5 antibodies stain areas of contact between corneal epithelium and basement membrane, indicating that rat laminin-5 from the medium is incorporated into basement membrane. We discuss the use of laminin-5 as a medium supplement for the culture of both epithelial cells and epithelial tissue explants.
大鼠细胞系804G能组装一种细胞外基质，这种基质不仅能诱导上皮细胞快速黏附与铺展，还能诱导一种名为半桥粒的细胞-基质附着装置的组装。这种基质的主要成分是层粘连蛋白-5。我们已从804G细胞条件培养液中纯化出大鼠层粘连蛋白-5。与补充了可溶性层粘连蛋白-5的培养液共同孵育的上皮细胞能快速黏附与铺展。此外，人类癌细胞在层粘连蛋白-5存在的情况下会发生显著的形态学变化，并形成类似上皮细胞片层的有序排列。可溶性大鼠层粘连蛋白-5在体外能被选择性地整合到上皮细胞的不溶性基质中，因为大鼠特异性层粘连蛋白-5抗体能对细胞-底物接触部位进行染色。向移植的人类角膜边缘添加含有可溶性层粘连蛋白-5的培养液可诱导半桥粒的组装，半桥粒是重要的细胞-基质附着装置。此外，大鼠特异性层粘连蛋白-5抗体能对角膜上皮与基底膜之间的接触区域进行染色，这表明培养液中的大鼠层粘连蛋白-5被整合到了基底膜中。我们讨论了层粘连蛋白-5作为上皮细胞和上皮组织外植体培养的培养液补充物的用途。
