Publications
Here is a selection of publications where different laminin isoforms were used to create more authentic cell culture systems.
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.
Dynamic blebbing: A bottleneck to human embryonic stem cell culture that can be overcome by Laminin-Integrin signaling
Weng J-H., Cheung C., Talbot P.Stem Cell Research, 2018
This study characterizes dynamic and apoptotic blebbing in human embryonic stem cells (hESC), identifies dynamic blebbing as a bottleneck to successful cell attachment during passaging, and demonstrates that dynamic blebbing can be rapidly stopped by plating cells on recombinant human laminin. In freshly plated hESC, dynamic and apoptotic blebbing differed in time of occurrence, bleb retraction rate, mitochondrial membrane potential, and caspase 3&7 activation. While dynamic blebbing can be controlled with drugs that inhibit myosin II, these methods have off-target effects and are not suitable for clinical applications. Recombinant human laminin-521 or addition of laminin-111 to Matrigel provided a safe method to drastically decrease dynamic blebbing and improve cell attachment with proteins normally found in the inner cell mass. Inhibition of focal adhesion kinase, which is activated by binding of integrins to laminin, prolonged dynamic blebbing and inhibited attachment. These data show that hESC binds rapidly to laminins through an integrin, which activates focal adhesion kinase that in turn downregulates dynamic blebbing. Laminins enabled hESC to rapidly attach during passaging, improved plating efficiency, enabled passaging of single pluripotent stem cells, and avoided the use of inhibitors that have non-specific off-target effects. These data provide a strategy for improving hESC culture using biologically safe recombinant human proteins.
Laminin-511 is an epithelial message promoting dermal papilla development and function during early hair morphogenesis
Jing Gao, Mindy C. DeRouen, Chih-Hsin Chen, Michael Nguyen, Ngon T. Nguyen, Hiroyuki Ido, Kenji Harada, Kiyotoshi Sekiguchi, Bruce A. Morgan, Jeffery H. Miner, Anthony E. Oro, and M. Peter Marinkovich. Genes Dev, 2008
Abnormal adhesion of grafted corneal endothelial cells (CECs) affects the application of intracameral injection for corneal endothelial dysfunction therapy. The authors explored whether laminin 511 (LN511) improves the therapeutic function of the intracameral CEC injection. Injected LN511 was found to be able to settle and form a coating on the posterior surface of Descemet's membrane (DM). The data suggests that the strategy of LN511 precoating and CECs' intracameral injection could be a potential method for the therapy of corneal endothelial dysfunction.
Laminin-10 is crucial for hair morphogenesis
Jie Li, Julia Tzu, Yi Chen, Yan-Ping Zhang, Ngon T Nguyen, Jing Gao, Maria Bradley, Douglas R Keene, Anthony E Oro, Jeffrey H Miner, M Peter Marinkovich. EMBO J, 2003
This article describes the essential role of laminin-10 (laminin-511) in hair follicle development.