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HOW TO CULTURE EPITHELIAL CELLS ON LAMININ SUBSTRATES
Epithelial tissues contain laminin-521, laminin-511 and laminin-332
Epithelial tissues are sheet of cells that line the surfaces and cavities of the whole body, including the skin, oral cavity, lung, gastrointestinal and urinary tracts, hair follicles and many glands. The epithelial cells are polarized with the basolateral surface resting on a basement membrane that contain high levels of alpha-5 laminins and laminin-332 (Domogatskaya, 2012). The expression level of the laminin-332 does not change significantly with age, whereas laminin-511 and laminin-521 appeared to decrease in the basement membrane underlying the epidermis in adult skin (Pouliot, 2002). Epithelial basement membranes also contain laminin-311 and to a lesser extent laminin-321 (Domogatskaya, 2012). Laminin-311 forms a covalent complex with laminin-332 in epithelial tissue (Champliaud, 1996) and has been shown to participate in mechanically induced signal transduction after mechanical stimulation (Jones, 2005).
Laminin is important for proper anchorage of epithelial cells
Laminin-332 supports the stable anchoring of basal keratinocytes to the epidermal basement membrane. In normal skin, stationary epithelial cells bind laminin-332 with high affinity for the α6β4 laminin–specific integrin as well as hemidesmosomes components (Ghohestani, 2001; Litjens, 2006; Nishiuchi, 2006). Laminin-332 binds tightly to the noncollagenous domain of type VII collagen of anchoring fibrils, important for proper anchorage of epithelial cells (Rousselle, 1997).
Mutations in the genes that encode the laminin-332 subunits, abrogate or perturb the functions of laminin-332. This may cause disruption of the epithelial α6β4 integrin/Laminin-332/type VII collagen adhesion complex which results in a skin blistering disease known as epidermolysis bullosa (Pulkkinen & Uitto, 1999; Vidal, 1995). A mutation specific for the shorter alpha-3 chain, α3A, likely to cause dysfunction of keratinocyte-mesenchymal communication induces cutaneous erosions, nail dystrophy, and exuberant vascular granulation tissue in certain epithelia (McLean, 2003).
Alpha-5 laminins are also expressed abundantly in the basement membrane underlying the epidermis and blood vessels in the dermis. Pouliot and colleagues demonstrated with in a vitro cell adhesion assays, that laminin-511 and laminin-521 are potent adhesive substrates for both neonatal and adult keratinocytes and that this adhesion is mediated by the α3β1 and α6β4 integrins (Pouliot, 2002).
Laminin promotes proliferation and migration of epithelial cells
LAMININ KEY ADVANTAGES
Defined and animal component-free cell culture matrix for clinical compliance.
A biologically relevant culture environment for epthelial cells
The epithelial cells are resting on a basement membrane that contain high levels of laminin-521, laminin-511 and laminin-332.
Laminin-511 and laminin-521 are potent adhesive substrates for both neonatal and adult keratinocytes and that this adhesion is mediated by the α3β1 and α6β4 integrins.
Laminin-332 promotes fast spreading of epithelial cells and functions as a motility factor for wound healing and cancer invasion.
No lot-to-lot variability for standardized experiments with less variation.
Scientifically validated in high-impact journals.