Publications
Here is a selection of publications where different laminin isoforms were used to create more authentic cell culture systems.
Laminin 411 mediates endothelial specification via multiple signaling axes that converge on β-catenin
Hall M.L., Givens S., Santosh N., Iacovino M., Kyba M. & Ogle B.M. Stem Cell Reports, 2022
The authors show that laminin-411 (Biolaminin 411) alone induces endothelial cell differentiation in human and mouse induced pluripotent stem cells over collagen I or Matrigel. Overall, the results indicate that the extracellular matrix contributes to endothelial differentiation through multiple avenues which converge at β-catenin. This protocol is an improvement to previous methods, excluding the use of undefined matrix substances and being less complex in terms of added small-molecule reagents.
Laminin 511 Precoating Promotes the Functional Recovery of Transplanted Corneal Endothelial Cells
Can Zhao, Qingjun Zhou, Haoyun Duan, Xin Wang, Yanni Jia, Yajie Gong, Wenjing Li, Chunxiao Dong, Zongyi Li, and Weiyun Shi. Tissue Eng Part A, 2020
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.
Glomerular endothelial cells and podocytes jointly synthesize laminin-1 and -11 chains
St John P.L. and Abrahamson D.R.Kidney International, 2001
During the glomerular basement membrane assembly, laminin-111 is initially expressed in vascular clefts of comma- ann S-shaped bodies and is eventually replaced by laminin-521 which persists into maturation. Post-fixation immunoelectron microscopy of developing mouse and 2-3 days old mice kidney was performed and showed Intracellular labeling for laminin-111 and laminin-521 in developing glomerular endothelial cells and podocytes. In early capillary loop stage, GBMs laminin-111 was absent but the expression of a5 was strong (developmental switch). The results show that both endothelial cells and podocytes synthesize laminin but the highest level of laminin synthesis by endothelial cells.
Switch in Laminin β2 to Laminin β1 Isoforms During Aging Controls Endothelial Cell Functions
Wagner J.U.G., Chavakis E., Rogg E.M, Muhly-Reinholz M., Glaser S.F., Günther S., John D., Bonini F, Zeiher A.M., Schaefer L., Hannocks M-J, Boon R.A., Dimmeler S.Arterioscler Thromb Vasc Biol, 2018
Here, the authors aim to decipher the role of the microenvironment underlying the impairment of endothelial cell functions by aging. RNA sequencing of isolated cardiac endothelial cells derived from young and 18-month-old mouse hearts revealed that aging affects the endothelial expression of genes encoding extracellular matrix proteins, specifically the laminin β1 (Lamb1) and laminin β2 (Lamb2) chains. Whereas Lamb1 was upregulated, Lamb2 was decreased in endothelial cells in old mice compared with young controls. A similar change in expression patterns was observed after induction of acute myocardial infarction. Mimicking aging and injury conditions by plating endothelial cells on laminin β1–containing laminin 411 matrix impaired endothelial cell adhesion, migration, and tube formation and augmented endothelial-to-mesenchymal transition and endothelial detachment compared with laminin 421, which contains the laminin β2 chain. Because laminins can signal via integrin receptors, the authors determined the activation of ITGB1 (integrin β1). Laminin 421 coating induced a higher activation of ITGB1 compared with laminin 411. siRNA-mediated silencing of ITGB1 reduced laminin β2–dependent adhesion, suggesting that laminin β2 more efficiently activates ITGB1. Mimicking age-related modulation of laminin β1 versus β2 chain expression changes the functional properties and phenotype of endothelial cells. The dysregulation of the extracellular matrix during vascular aging may contribute to an age-associated impairment of organ function and fibrosis.
Differentiation of Human Embryonic Stem Cells to Endothelial Progenitor Cells on Laminins in Defined and Xeno-free Systems
Nguyen M.T.X., Okina E., Chai X., Tan K.H., Hovatta O., Ghosh S., Tryggvason K. Stem Cell Reports, 2016
Here, the authors developed a chemically defined, xeno-free protocol for differentiation of hESCs to endothelial progenitor cells (EPCs) using LN521 as the main culture substrate. The EPCs derived are functional and expressed both progenitor and mature endothelial markers. The authors were able to generate about 95% functional EPCs defined as VEGFR2+CD34+CD31+VE-Cadherin+. RNA-sequencing analyses of hESCs, EPCs, and primary human umbilical vein endothelial cells show differentiation-related EC expression signatures, regarding basement membrane composition, cell-matrix interactions, and changes in endothelial lineage markers. Six-week continuous culturing allows the hESC derived EPSs to mature further, relative to HUVECs. These results may facilitate the production of stable ECs for the treatment of vascular diseases and in vitro cell modeling.
Laminin isoforms in endothelial and perivascular basement membranes
Yuosif L., Russo J.D., Sorokin L.Cell Adh Migr., 2013
α4 and α5 chain laminins are the predominant isoforms in the basal lamina of vascular endothelial cells. While the α4 chain is expressed ubiquitously throughout different developmental stages, the prominent expression of α5 chain appears postnatally and its distribution varies with vessel type.
3D map of the human corneal endothelial cell
He Z., Forest F., Gain P., Rageade D., Bernard A., Acquart S., Peoch M., Defoe D.M., Thuret G. Scientific reports, 2016
Human corneal endothelial cells (CECs) are highly polarized flat cells that separate the cornea from the aqueous humor. Their apical surface, in contact with aqueous humor is hexagonal, whereas their basal surface is irregular. Here, the authors characterized the structure of human CECs in 3D using confocal microscopy of immunostained whole corneas in which cells and their interrelationships remain intact. Hexagonality of the apical surface was maintained by the interaction between tight junctions and a submembraneous network of actomyosin. Lateral membranes presented complex expansions resembling interdigitated foot processes at the basal surface. Integrin α3β1 was the only protein found exclusively at the basal surface, forming an almost homogenous layer that follows the slightly bumpy surface of Descemet’s membrane. Ligands of integrin α3β1, such as laminin-332, laminin-511, and laminin-521 constitute efficient coating substances that improve the yield of in vitro CEC cultures. This first 3D map aids our understanding of the morphologic and functional specificity of CECs and could be used as a reference for characterizing future cell therapy products destined to treat endothelial dysfunctions.
Endothelial basement membrane limits tip cell formation by inducing Dll4/Notch signalling in vivoThe extracellular matrix protein laminin-10 promotes blood–brain barrier repair after hypoxia and inflammation in vitro
Kangwantas K., Pinteaux E., Penny J.Journal of neuroinflammation 2016
Integrity of the BBB is primarily maintained by brain endothelial cells, the tight junctions between them and their attachment to the blood vessel basement membrane (mainly composed fibronectin, collagen IV, and laminin-411 and -511). Here the authors used an in vitro model of the BBB, composed of primary rat brain endothelial cells grown on these different ECM proteins. The in vitro BBB model was exposed to oxygen-glucose deprivation with or without reoxygenation, and in the absence or the presence of IL-1β in order to mimic the ischemic and inflammatory conditions that occur during stroke. They show that laminin-511 plays a key role in maintenance of BBB integrity and that it’s a key ECM molecule involved in BBB repair after hypoxic injury and inflammation. The brain endothelial cells did not adhere well to laminin-411.
Endothelial Cell Laminin Isoforms, Laminins 8 and 10, Play Decisive Roles in T Cell Recruitment Across the Blood–Brain Barrier in Experimental Autoimmune Encephalomyelitis
Sixt M., Engelhardt B., Pausch F., Hallmann R., Wendler O., Sorokin L.M. J Cell Biol., 2001
Laminin-411 and laminin-511 are described as the major laminin isoforms in vascular basement membranes. Their expression was influenced by pro-inflammatory cytokines or angiostatic agents. Inflammatory cuffs occurred exclusively around endothelial basement membranes containing laminin-411, whereas in the presence of laminin-511 no infiltration was detectable. Integrin α6 and β-dystroglycan were prominent in CNS blood vessels, whereas no staining was observed for integrin α3, α7, and β4 subunits. One of the major laminin receptors, integrin α6β1, was localized predominantly on the endothelial cells, where it is likely to mediate interactions with the endothelial cell laminin-411 and -511, whereas astrocyte endfeet appear to utilize a different receptor for interactions with the parenchymal laminins. β-Dystroglycan occurred predominantly on astrocyte endfeet.
Laminin-421 produced by lymphatic endothelial cells induces chemotaxis for human melanoma cells
Saito N., Hamada J., Furukawa H., Tsutsumida A., Oyama A., Funayama E., Saito A., Tsuji T., Tada M., Moriuchi T., Yamamoto Y.Pigment Cell Melanoma Res., 2009
Here, the authors investigate the molecular mechanism of lymphatic metastasis. They examined the influence of interactions between normal lymphatic endothelial cells (LECs) and melanoma cells on cell migration. LEC conditioned medium (LEC-CM) contained chemotactic and chemokinetic activities for human melanoma cell lines. The chemotactic activity of LEC-CM was abolished by immunodepletion with anti-laminin-1 antibody. And immunoprecipitation and Western blot analyses revealed that LEC-CM contained laminin a4 and 5, b1 and 2, and c1, corresponding to isoforms -521, -511, -421 and -411. When melanoma C8161 cells were treated with function-blocking antibodies to integrin a3 or a6, their chemotactic responses to LEC-CM were markedly reduced. Furthermore, the knock-down of tetraspanin CD151 weakened the chemotactic responses of C8161 and MeWo cells to LEC-CM. These data suggest that laminin secreted by LEC possibly facilitates lymphatic metastasis through the induction of chemotaxis of melanoma cells.