Publications
Here is a selection of publications where different laminin isoforms were used to create more authentic cell culture systems.
Laminin-521 Promotes Rat Bone Marrow Mesenchymal Stem Cell Sheet Formation on Light-induced Cell Sheet Technology
Jiang Z., Xi Y., Lai K., Wang Y., Wang H., Yang G.BioMed Research International, 2016
The cell sheet technology is an area of research that is of great interest for tissue engineering and regenerative medicine. Here, the authors investigated the effects of an ECM coating on rat bone marrow mesenchymal stem cells (rBMSC) cultured on light-induced TiO2 nanodot films. Cell sheets can be detached on a TiO2 nanodot-coated quartz substrate by using UV365 illumination. The effects of rat fibronectin, human recombinant laminin-521, -511, -421, and -111 on the formation of cell sheets were investigated and also compared to uncoated films. The result showed the highest success for laminin-521. rBMSCs rapidly attached and spread on films coated with laminin-521 (1.2 ug/ml) and formed intact cell sheets after 5 days of culture. Laminin-521 promotes the formation of rBMSC sheets with good viability under hyperconfluent conditions (4 to 8 layers of cells). The cells also maintained multilineage potential, including osteogenic, adipogenic, and chondrogenic differentiation. The cell sheets formed had rich ECM (including collagen I) and cells were connected with each other in dense network-like tissue. rBMSC cultured on uncoated surface and cells partially detached and failed to form cell sheets. In summary, laminin-521 and UV365 illumination systems provided a simple, rapid, and effective cell sheets strategy.
Laminin isoforms and their receptors in the developing kidney
Ekblom P., Klein G., Ekblom M., Sorokin L.Am J Kidney Dis., 1991
Epithelial cells have a polarized morphology, with distinct basal, lateral, and apical cell surfaces. It would be of considerable interest to know how the polarized morphology develops during embryogenesis. Both the tubular and glomerular epithelial cells of the kidney develop from mesenchymal stem cells during embryogenesis. Unique conversion of nonpolar cells to polarized epithelial cells thus occurs in the embryonic kidney. This conversion also occurs in vitro if the mesenchymal cells are properly induced. Organ cultures of mesenchymal cells from the mouse embryonic kidney have therefore been much used to study the development of epithelial cell polarity. We have used this model system to study the role of basement membrane glycoproteins in development. The results obtained suggest that laminins are particularly important for epithelial cell development. There are many different types of laminins. Developing kidney tubule cells synthesize that promote development by interacting with specific integrin receptors on the cell surface. The mesenchymal stem cells also produce laminin.
Abnormal Wnt and PI3Kinase Signaling in the Malformed Intestine of lama5 Deficient Mice
Ritié L., Spenlé C., Lacroute J.I., Bolcato-Bellemin A-L., Lefebvre O., Bole-Feysot C., Jost B., Klein A., Arnold C., Kedinger M., Bagnard D., Orend G., Simon-Assmann P. PLoS One, 2012
Laminin-511 is highly expressed in the intestine. To understand the mechanistic role of laminin-511 in tissue homeostasis, the researchers used RNA profiling of embryonic intestinal tissue of lama5 knockout mice and identified a lama5 specific gene expression signature. They show that laminin a5 plays a crucial role in both epithelial and mesenchymal (smooth muscle) cell behavior by inhibiting Wnt and activating PI3K signaling. We conclude that conflicting signals are elicited in the absence of lama5, which alter cell adhesion, migration as well as epithelial and muscle differentiation. The LMa5 deficient intestine also displays a smooth muscle defect and myogenic differentiation markers are affected. Laminin-511 supports adhesion of epithelial cells and Akt phosphorylation. Laminin-511 stimulates the spreading of epithelial and muscle cells (compared to laminin-111). Inhibition of Akt with wortmannin abolished spreading of epithelial cells on laminin-511 as evidenced by cell laminin-511 specifically activates Akt through the PI3K pathway in intestinal epithelial but not in mesenchymal cells. Cell migration was also higher on Laminin-511. Laminin-511 also protects cells against H2O2-induced apoptosis.
Laminins in the Developing and Adult Human Small Intestine: Relation With the Functional Absorptive Unit
Teller I.C., Auclair J., Herring E., Gauthier R., Ménard D., Beaulieu J-F.Developmental dynamics, 2007
Here, the expression of the five laminin a-chains was analyzed in the developing and mature human small intestine at the protein and transcript levels in order to further delineate specific involvement of individual laminins in relation to the epithelial cell state as defined along the functional crypt-villus axis. The results show that all of the a-laminins are expressed in significant amounts in the small intestine relative to a panel of other tissues and organs. Distinct epithelial and mesenchymal origins, as well as a differential occurrence in intestinal basement membranes according to developmental stage, along the crypt-villus axis and in compartment-related experimental intestinal cell models. Taken together, the data point out the prime importance of a2-, a3-, and a5-containing laminins for the development and maintenance of the functional human intestinal epithelium.
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.
Monoclonal antibodies to human laminin α4 chain globular domain inhibit tumor cell adhesion and migration on laminins 411 and 421, and binding of α6β1 integrin and MCAM to α4-laminins
Ishikawa T., Wondimu Z., Oikawa Y., Ingerpuu S., Virtanen I., Patarroyo M.Matrix Biology, 2014
α4-Laminins, such as laminins -411 and -421, are mesenchymal laminins expressed by vascular and lymphatic endothelial cells, leukocytes and other normal cell types. These laminins are recognized by α6β1 and α6β4 integrins and MCAM (CD146) and promote adhesion and migration of the cells. α4-Laminins are also expressed and secreted by some tumor cells and strongly promote tumor cell migration. Moreover, the abluminal side of blood and/or lymphatic vessels and the nerve perineurium, common tracks of tumor cell dissemination, express α4-laminins, and these laminin isoforms, when expressed in the stroma, may contribute to tumor invasion. In the present study, we examined ten mAbs to human laminin α4 chain for their reactivity with the isolated laminin α4 globular domain, their ability to inhibit tumor cell adhesion and migration on laminin-411 and -421, and their effect on the binding of α6β1 integrin and MCAM to both α4-laminins. The results indicate that mAbs to the laminin α4 globular domain are able to inhibit tumor cell adhesion and migration on laminin-411 and -421 and that α6β1 integrin and MCAM bind α4-laminins at very close sites on the globular domain. These reagents contribute to a better understanding of the biology of α4-laminins and may have a therapeutic potential in malignant and inflammatory diseases.
The Different Binding Properties of Cultured Human Corneal Endothelial Cell Subpopulations to Descemet’s Membrane Components
Toda M., Ueno M., Yamada J., Hiraga A., Tanaka H., Schlötzer-Schrehardt U., Sotozono C., Kinoshita S., Hamuro J.Invest Ophthalmol Vis Sci. 2016
In culture, human corneal endothelial cell (cHCEC) tend to enter into cell-state transition (CST), such as epithelial-to-mesenchymal transition (EMT) or fibrosis, thus resulting in the production of different subpopulations. In this study, the authors examined the binding ability of cHCECs subpopulations to major Descemet’s membrane components that distribute to the endothelial face; that is, laminin-511, -411, Type-IV collagen, and proteoglycans. Each subpopulation was prepared by controlling the culture conditions or by using magnetic cell separation and then confirmed by staining with several cell-surface markers. Binding abilities of HCEC subpopulations were examined by adding the cells to culture plates immobilized with collagens, laminins, or proteoglycans, and then centrifuging the plates. The cHCECs showed the best attachment to laminin laminin-521 and -511. The cells showed a weaker binding to laminin-411, laminin-332, Type-IV collagen. The minimum concentrations necessary for the observed cell binding in this study were as follows: laminin-521 and -511, 3 ng/mL; laminin-411, 2.85 ug/mL; Type-IV collagen, 250 ng/mL. Cells suspended in Opti-MEM-I or Opeguard-MA were bound to laminin, yet no binding was observed in cells suspended in BSS-Plus. Both the fully differentiated, mature cHCEC subpopulations and the epithelial-to-mesenchymal– transitioned (EMT)-phenotype subpopulation were found to attach to laminin- or collagen-coated plates. Interestingly, the binding properties to laminins differed among those subpopulations. Although the level of cells adhered to the laminin-411–coated plate was the same among the cHCEC subpopulations, the fully differentiated, mature cHCEC subpopulations were significantly more tightly bound to laminin-511 than was the EMT-phenotype subpopulations. These findings suggest that the binding ability of cHCECs to major Descemet’s membrane components is distinct among cHCEC subpopulations and that Opti-MEM-I and Opeguard-MA are useful cell-suspension vehicles for cell-injection therapy. This research group focused on developing a novel medical approach, termed cell-injection therapy, for the treatment of patients with endothelial dysfunction.
Superior Red Blood Cell Generation from Human Pluripotent Stem Cells Through a Novel Microcarrier-Based Embryoid Body Platform
Sivalingam J., Lam A.T., Chen H.Y., Yang B.X., Chen A.K., Reuveny S., Loh Y.H, Oh S.K. Tissue Eng Part C Methods, 2016
Conventional methods for hematopoietic differentiation of human pluripotent stem cells (hPSC) rely on embryoid body (EB) formation and/or co-culture with xenogeneic cell lines. In this study, the authors describe the development of a scalable, serum-free, xeno-free, and chemically defined microcarrier-based platform using human recombinant laminin-521 as an extracellular matrix (ECM) for hPSC expansion, EB formation, and subsequently hematopoietic differentiation of hPSC to red blood cells (RBS). Improved survival and better quality EBs generated with the microcarrier-based method resulted in significantly improved mesoderm induction and, when combined with hematopoietic differentiation, resulted in at least a 6-fold improvement in hematopoietic precursor expansion, potentially culminating in an 80-fold improvement in the yield of RBS generation compared to a conventional EB-based differentiation method. In addition, they show efficient terminal maturation and generation of mature enucleated RBCs using a co-culture system that comprised primary human mesenchymal stromal cells.
Laminin-521 promotes quiescence in isolated stellate cells from rat liver
Rohn F., Kordes C., Castoldi M., Götze S., Poschmann G., Stühler K., Herebian D., Benk A.S., Geiger F., Zhang T. Spatz J.P., Häussinger D.Biomaterials, 2018
Here, the authors show that Biolaminin 521 supports the quiescent state of HSC and that laminin a5 can be regarded as an important element of their niche in the space of Dissé. Hepatic stellate cells (HSC) are liver-resident mesenchymal stem cells, which reside in a quiescent state on a basement membrane-like structure in the space of Dissé. In the present study, a laminin a5 chain was detected in the space of Dissé of normal rat liver. Since HSC is critical for liver regeneration and can contribute to fibrosis in chronic liver diseases, the effect of laminins on HSC maintenance was investigated. Isolated rat HSC were seeded on uncoated polystyrene (PS) plates or PS plates coated with either laminin-521 (PS/LN-521) or laminin-211 (PS/LN-211). PS/LN-521 improved HSC adhesion and better preserved their retinoid stores as well as quiescence- and stem cell-associated phenotype, whereas HSC on PS/LN-211 or PS developed into myofibroblasts-like cells. To improve the homogeneity, as well as the presentation of laminin molecules on the culture surface to HSC, laminin-functionalized, gold-nanostructured glass surfaces, were generated. This approach further enhanced the expression of quiescence-associated genes in HSC.