Publications
Here is a selection of publications where different laminin isoforms were used to create more authentic cell culture systems.
Laminin-511 and integrin beta-1 in hair follicle development and basal cell carcinoma formation
DeRouen M.C., Zhen H., Tan S.H., Williams S., Marinkovich M.P., Oro A.E.BMC Dev Biol., 2010
With the use of a basal cell carcinoma (BCC) model system and mouse mutants the authors re-evaluate the role of laminin-511 in epithelial invagination in the skin. Blocking laminin 511 and 332 in BCCs maintains primary cilia and Shh signalling, but prevents invagination. Similarly, in laminin-511 and dermal beta-1 integrin mutants, dermal papilla development and primary cilia formation are normal. Dermal beta-1 integrin mutants have normal hair follicle development. They conclude that laminin-511 has a primary role of promoting hair follicle epithelial downgrowth without affecting dermal primary cilia and Shh target gene induction.
Spatial and temporal control of laminin-332 (5) and -511 (10) expression during induction of anagen hair growth
Koji Sugawara, Daisuke Tsuruta, Hiromi Kobayashi, Kazuo Ikeda, Susan B Hopkinson, Jonathan C R Jones, Masamitsu Ishii. J Histochem Cytochem, 2007
The results in this article indicate a positive role for laminin-511 and a negative role for laminin-332 on hair growth.
FK506 positively regulates the migratory potential of melanocyte-derived cells by enhancing syndecan-2 expression
Hyejung Jung and Eok-Soo Oh Pigment Cell Melanoma Res, 2016
The authors report that topical tacrolimus (FK506) treatment, known to promote repigmentation, enhanced cell spreading on laminin-332 and increased migration in both melanocytes and melanoma cells by enhancing syndecan-2 expression.
Laminin 10/11: an alternative adhesive ligand for epidermal keratinocytes with a functional role in promoting proliferation and migration
Pouliot N., Saunders N.A., Kaur P. Exp Dermatol., 2002
The authors investigated the expression and function of laminin-511 and -521 in neonatal and adult human skin. They found that the laminin-a5 chain is expressed abundantly in the basement membrane underlying the interfollicular epidermis and the blood vessels in the dermis. Interestingly, while the expression level of the well-studied laminin-5 isoform did not change significantly with age, laminin-511 and -521 appeared to decrease in the basement membrane underlying the epidermis, in adult skin. In contrast, the levels of laminin-511 and -521 in the basement membrane underlying blood vessels remained unchanged in neonatal vs. adult skin.
Development of a biomaterial associated with mesenchymal stem cells and keratinocytes for use as a skin substitute
Steffens D., Mathor M.B., Santi B.T.S., Luco D.P., Pranke P.Regen. Med., 2015
Here they developed s scaffolds of poly-DL-lactic acid with and without the linkage of laminin-332, bringing together MSCs and keratinocytes aimed for treatment as a new skin substitute. Three groups of scaffolds were studied: 1) poly-DL-lactic acid (PDLLA), 2) hydrolyzed PDLLA (PDLLA/NaOH) and 3) PDLLA/Lam which is a PDLLA/NaOH scaffold linked to laminin-332. The results corroborate the hypothesis that laminin influenced the adhesion of the MSCs. Laminin significantly promoted the adhesion and spreading of proliferating oral and epidermal keratinocytes compared with collagen nanofibers only. The use of biocompatible and biodegradable polymers associated with the properties of laminin leads to an improvement in the adherence and viability of the cells, showing LN-332 is beneficial for the growth of MSCs and keratinocytes.
Polymerized Laminin-332 Matrix Supports Rapid and Tight Adhesion of Keratinocytes, Suppressing Cell Migration
Kariya Y., Sato H., Katou N., Kariya Y., Miyazaki K.PLOS ONE, 2012
Laminin-332 is known to supports the stable anchoring of basal keratinocytes to the epidermal basement membrane but is also a motility factor for wound healing and cancer invasion. Here they investigated Laminin-332 matrices deposited by normal human keratinocytes and many cancer cell lines. All types of cells efficiently deposited Laminin-332 on the culture plates in specific patterns. On the contrary, laminins containing laminin ß1 and/or c1 chains (such as Lm511 and Lm311) were not deposited on the culture plates even if secreted into the culture medium. The deposited Laminin-332 matrix showed a mesh-like network structure as analyzed by electron microscopy, suggesting that Lm332 was highly polymerized. Laminin-332 matrix rather suppressed the migration of keratinocytes as compared with purified Lm332 (not a BioLamina product), which highly promoted cell migration. The Lm332 matrix support adhesion of keratinocytes much more strongly and stably than purified Lm332. Integrin a3ß1 bound to the Lm332 matrix at a three times higher level than purified Lm332. These results indicate that the polymerized Lm332 matrix supports stable cell adhesion whereas unassembled soluble Lm332 supports cell migration. The question is though how the purified Ln-332 looked like. Difficult to purify and might be fractionated.
Laminin heparin-binding peptides bind to several growth factors and enhance diabetic wound healing
Ishihara J., Ishihara A., Fukunaga K., Sasaki K., White M.J.V., Briquez P.S., Hubbell J.A.Nature Communications, 2018
In this article, the authors show that multiple laminin isoforms promiscuously bind to growth factors with high affinity, through their heparin-binding domains located in the α chain laminin-type G (LG) domains. These domains also bind to syndecan cell-surface receptors, promoting attachment of fibroblasts and endothelial cells. The authors explore the application of these multifunctional laminin HBDs in wound healing in the type-2 diabetic mouse and demonstrate that covalent incorporation of laminin HBDs into fibrin matrices improve retention of GFs and significantly enhances the efficacy of vascular endothelial cell growth factor (VEGF-A165) and platelet-derived growth factor (PDGF-BB) in promoting wound healing in vivo, under conditions where the GFs alone in fibrin are inefficacious.
Biologically relevant laminin as a chemically defined and fully human platform for human epidermal keratinocyte culture
Tjin M.S., Chua A.W.C, Moreno-Moral A., Chong L.Y. , Tang P.Y., Harmston N.P., Cai Z., Petretto E., Tan B.K., Tryggvason K. Nature Communications, 2018
The current expansion of autologous human keratinocytes to resurface severe wound defects still relies on murine feeder layer and calf serum in the cell culture system. Here, the authors report a completely xeno-free and defined culture system utilizing skin-specific laminin cell culture substrates which enables robust expansion of adult human skin keratinocytes. The authors characterized the human skin basement membrane and murine feeder layer 3T3- J2 and have identified two biologically relevant recombinant laminins, laminin 511 and 421, as potential candidates to replace the murine feeder. The authors report a completely xeno-free and defined culture system utilizing these laminins which enables robust expansion of adult human skin keratinocytes, comparable to the 3T3-J2 co-culture system in terms of basal markers’ profile, colony-forming efficiency and the ability to form a normal stratified epidermal structure in both in vitro and in vivo models. Human keratinocytes cultured either on laminin 511 or 421 were able to generate a fully stratified epidermal layer in vivo similar to that on the 3T3 co-culture system. The results show that the proposed system may not only provide safer keratinocyte use in the clinics but also facilitate the broader use of other cultured human epithelial cells in regenerative medicine.
Chemically defined and xenogeneic-free culture method for human epidermal keratinocytes on laminin-based matrices
Tjin M.S., Chua A.W.C, Tryggvason K. Nature Protocols, 2020
In this protocol, the authors describe how to implement a defined, xeno-free culture system that supports long-term ex vivo expansion of functional human epidermal keratinocytes. Laminins, skin-specific basement membrane proteins play important roles in the maintenance of phenotypic integrity and in supporting the survival of keratinocytes that are adhered to them. The fully human keratinocyte culture system is presented in this article is ‘regulatory friendly’ and increases the potential of epithelial cellular therapy, which can be expanded to treat less severe burns and other skin defects, such as chronic diabetic wounds. It takes between 7 and 14 d to obtain an initial culture and a secondary culture from the primary culture can be expanded up to 20-fold within 4–5 d once cells reach confluency.
Decrease of laminin‑511 in the basement membrane due to photoaging reduces epidermal stem/progenitor cells
Iriyama S., Yasuda M., Nishikawa S., Takai E., Hosoi J., Amano S.Scientific reports, 2020
Here, the authors examine how photoaging affects the function of Inter-follicular epidermal stem cells (IFE-SCs) which regulate epidermal proliferation and differentiation. It is known that daily sunlight disrupts epidermal homeostasis and the authors found that sun-exposed skin showed a decrease of MCSP-positive and β1-integrin-positive cells concomitantly with a decrease of laminin-511 at the dermal-epidermal junction (DEJ), as compared with sun-protected skin. Higher levels of laminin-511 were associated with increased colony formation efficiency, higher expression levels of MCSP as well as other stem cell markers in human skin. UVB exposure to cultured human skin impaired laminin-511 integrity at the dermal-epidermal junction and reduced MCSP-positive basal epidermal cells as well as K15-positive cells. Combined treatment with matrix metalloproteinase and heparanase inhibitors protected the integrity of laminin-511 and inhibited the reduction of MCSP-positive cells and K15-positive cells. These results suggest that photoaging may reduce the levels of MCSP-positive and K15-positive epidermal stem/ progenitor cells in the epidermis via loss of laminin-511 at the dermal-epidermal junction.