Publications
Here is a selection of publications where different laminin isoforms were used to create more authentic cell culture systems.
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.
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.
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.
Laminin 521 stabilizes the pluripotency expression pattern of human embryonic stem cells initially derived on feeder cells
Albalushi H., Kurek M., Karlsson L., Landreh L., Rós Kjartansdóttir K., Söder O., Hovatta O., Stukenborg J-B.Stem Cell International, 2017
In this article, the authors tested the effect of the laminin-521 substrate on cultured hES cells. Five male hES cell lines, originally derived on human foreskin fibroblasts (hFFs), were cultured on hFF, Matrigel, laminin-521 or laminin-121 for nine passages. Variations in gene expression related to pluripotency, stemness, and male germ and somatic gonadal cells at different passages and different culture methods were evaluated. All cell lines expressed pluripotency markers at protein and gene level and were able to differentiate into cell types of the three germ layers after being cultured on laminin-521 for nine passages. Laminin 521 had no obvious effect on the expression of genes related to male germ cells and somatic gonadal cells when used as a matrix for hES cell cultures. Importantly, reduction in variation of pluripotency marker expression was observed after culturing the cells on laminin-521 compared to culture on hFFs, and could be reduced further with an increased culture period on laminin-521. Changing the cell culture medium of hES cells on hFFs from Dulbecco's Modified Eagle Medium (DMEM) to NutriStem, which is commonly used for culturing the cells on matrices, did not affect the variation of pluripotency genes and genes related to stemness expression. hES cells cultured on laminin-521 were more homogenous, attached better and grew faster compared to hES cells cultured on matrigel. The results show that laminin-521 provides optimal culture conditions for adaptation to feeder and xeno-free conditions of hES cells derived and cultured on feeder cells. Moreover, laminin-521 has a positive effect on stabilizing and homogenizing pluripotent gene expression profiles between hES cell lines provides the first step towards more controllable and robust culture conditions for hES cells.