Here is a selection of publications where different laminin isoforms were used to create more authentic cell culture systems.

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  • Laminin a4 Deficient Mice Exhibit Decreased Capacity for Adipose Tissue Expansion and Weight Gain

    Vaicik M.K., Thyboll Kortesmaa J., Movérare-Skrtic S., Kortesmaa J., Soininen R., Bergström G., Ohlsson C., Chong L.Y., Rozell B., Emont M., Cohen R.N., Brey E.M., Tryggvason K.PLOS ONE, 2014

    Staining was performed for known adipose tissue BM proteins. In wild-type control mice the a2 and a4 chains of laminin were present in the BM surrounding mature adipocytes. Laminin a5 was not observed in mouse adipocyte BM. In this manuscript, we describe the role of laminin a4, a specialized ECM protein surrounding adipocytes, on weight gain and adipose tissue function. Adipose tissue accumulation, lipogenesis, and structure were examined in mice with a null mutation of the laminin a4 gene (Lama42/2) and compared to wild-type (Lama4+/+) control animals. Lama42/2 mice exhibited reduced weight gain in response to both age and high-fat diet. Interestingly, the mice had decreased adipose tissue mass and altered lipogenesis in a depot-specific manner. In particular, epididymal adipose tissue mass was specifically decreased in knock-out mice, and there was also a defect in lipogenesis in this depot as well. In contrast, no such differences were observed in subcutaneous adipose tissue at 14 weeks. The results suggest that laminin a4 influences adipose tissue structure and function in a depot-specific manner.

  • Roles for Laminin in Embryogenesis: Exencephaly, Syndactyly, and Placentopathy in Mice Lacking the Laminin α5 Chain

    Miner J.H., Cunningham J., Sanes J.R. J Cell Biol., 1998

    Here, we show that the laminin α5 chain is required during embryogenesis. The α5 chain is present in virtually all BLs of early somite stage embryos and then becomes restricted to specific BLs as development proceeds. BLs that lose α5 retain or acquire other α chains. Embryos lacking laminin α5 die late in embryogenesis. They exhibit multiple developmental defects, including failure of anterior neural tube closure. In addition to exencephaly, syndactyly, and placentopathy, several defects were seen in internal organs of Lama5 −/− embryos, including small or absent kidneys. Some of these defects were variable in severity and penetrance, but all can be associated with BLs that normally contain laminin α5.

  • Laminin a4-Null Mutant Mice Develop Chronic Kidney Disease with Persistent Overexpression of Platelet-Derived Growth Factor

    Abrass C.K., Hansen K.M., Patton B.L.Matrix Pathobiology, 2010

    Laminin-8/9 is synthesized by rat glomerular mesangial cells and is required for PDGF-induced mesangial cell migration. Here the authors show that Lama4-/- mice have progressive glomerular and tubulointerstitial fibrosis. These mice have a significant increase in the expression of platelet-derived growth factor (PDGF)-BB, PDGF-DD, and PDGF receptor beta in association with immature glomerular and peritubular capillaries. In addition, mesangial cell exposure to alpha4-containing laminins results in the down-regulation of PDGF receptor mRNA and protein, suggesting a direct effect of LN411/LN421 on vessel maturation. These data suggest that the failure of laminin alpha4-mediated down-regulation of PDGF activity contributes to the progressive renal lesions in this animal model. 

  • 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.

  • Culturing functional pancreatic islets on α5-laminins and curative transplantation to diabetic mice

    Sigmundsson K., Ojala J.R.M., Öhman M.K. Österholm A-M., Moreno-Moral A., Domogatskaya A., Yen Chong L., Sun Y., Chai X., Steele J.A.M, George B., Patarroyo M., Nilsson A-S., Rodin S., Ghosh S., Stevens M.M., Petretto E., Tryggvason K. Matrix Biology, 2018

    Here, the authors have developed a novel method to grow and maintain normoxic and functional islets which may significantly enhance the efficacy of islet transplantation treatment for diabetes. A key component of this method is the coating with biologically relevant laminins, found in the peri-islet capsule and BM of islet capillaries. Islets cultured in vitro on α5-laminins adhere and spread to form layers of 1-3 cells in thickness while maintaining cell-to-cell contacts. The cells remained normoxic and functional for at least 7 days in culture. In contrast, spherical islets kept in suspension developed hypoxia and central necrosis within 16 hours. Mouse islets plated on laminin-521 could be cultured in a serum-free mTeSR1 medium for an extended period of time. Approximately 20% of islet cells showed a co-expression of insulin and glucagon. The double-hormone expression was confirmed in histological analyses of mouse and monkey pancreata. The flattened islets start robust cell proliferation after a lag period of approximately two weeks in a serum-free mTeSR1 culture on laminin-521. Transplantation mouse islets cultured on α5-laminin-coated polydimethylsiloxane membranes for 3–7 days normalized blood glucose already within 3 days in mice with streptozotocin-induced diabetes.

  • Laminin 511 and WNT signalling sustain prolonged expansion of hiPSC-derived hippocampal progenitors

    Keagan Dunville, Fabrizio Tonelli, Elena Novelli, Azzurra Codino, Verediana Massa, Anna Maria Frontino, Silvia Galfrè, Francesca Biondi, Stefano Gustincich, Matteo Caleo , Luca Pandolfini, Claudia Alia, and Federico Cremisi. Development, 2022

    The authors identify laminin-511 as a crucial laminin isoform for prolonging the neural stem cell (NSC) state and extending hippocampal NSC proliferation for over 200 days in vitro. Biolaminin 511 supported adhesion and cell cycle progression of the dividing hippocampal progenitors. LN511 was crucial in supporting progenitor proliferation, inhibiting differentiation, and sustaining a gene expression profile responsible for maintaining a hippocampal neurogenic niche for extended periods compared with isoforms LN121, LN332, LN441, and with a mouse laminin product. The study involved a novel protocol for differentiating hippocampal NPCs from human induced pluripotent stem cells via a WNT actuator. The differentiation capability of both young and older NPC populations was retained when tested by xenografting into mice.

  • Integrin-dependent response to laminin-511 regulates breast tumor cell invasion and metastasis

    Kusuma N., Denoyer D., Eble J.A., Redvers R.P., Parker B.S., Pelzer R. Anderson R.L., Pouliot N.International Journal of Cancer, 2011

    Laminin-511 is a potent adhesive and migratory substrate for metastatic breast tumor cells in vitro and its expression correlates with tumor grade and metastatic potential in vivo. Here the authors compared the metastatic potential of 4T1 mammary carcinoma cells to that of 4T1 variants isolated by repeated chemotactic migration toward LM-511 in vitro (4T1LMF4) followed by serial injection into the mammary gland and recovery of spontaneous metastases from the bone (4T1BM2). Variant subpopulations exhibited a distinct morphology on LM-511 and increased expression of b1 and b4 integrins compared to parental 4T1 cells. Importantly, mice inoculated with 4T1LMF4 and 4T1BM2 variants showed a 2.5- to 4-fold increase in the incidence of spontaneous metastasis to bone compared to 4T1 tumor-bearing mice. Functionally, 4T1BM2 variants were more adherent and more invasive toward LM-511 than parental 4T1 cells. Treatment of 4T1BM2 cells with lebein-1, a dis-integrin with selectivity toward LM-type integrin receptors, potently inhibited their migration and invasion toward LM-511. Similarly, a3b1 integrin-dependent migration and invasion of human MDA-MB-231 breast carcinoma cells toward LM-511 were significantly inhibited by lebein-1. Taken together, these results provide strong evidence that LM-511 contributes to the metastasis of breast tumors and suggest that targeting integrin-LM- 511 interactions with lebein-1 or other inhibitors of LM-511 receptors may have therapeutic potential for patients with advanced breast cancer.

  • Laminin-211 controls thymocyte—thymic epithelial cell interactions

    Laminin-211 controls thymocyte—thymic epithelial cell interactions

    Ocampo J.S.P., de Brito J.M,  Corrêa-de-Santana E., Borojevic R., Serra Villa-Verde D.M., Savino W.
    Cellular Immunology, 2008

    Thymocyte differentiation occurs within the thymic microenvironment. Previous experiments showed that laminin mediates interactions between thymocytes and thymic epithelial cells (TEC) in mice. Here, the authors show constitutive gene expression of various laminin chains in TEC preparations, comprising laminin-111 and laminin-211 isoforms. Immunocytochemistry revealed a selective laminin-211 distribution in the thymic lobules. In vitro, functional assays revealed that laminin-211 enhances TEC/thymocyte adhesion and thymocyte release from thymic nurse cells, as well as the reconstitution of these complexes. Conversely, these interactions are blocked by monoclonal antibodies specific for laminin-211 and the laminin receptor VLA-6. The results show that distinct laminin isoforms in the human thymus are relevant for lymphoepithelial interactions.

  • Laminins affect T cell trafficking and allograft fate

    Warren K.J., Iwami D., Harris D.G, Bromberg J.S., Burrell B.E.The Journal of Clinical Investigation, 2014

    Lymph nodes (LNs) are integral sites for the generation of immune tolerance and migration of CD4+ T cells, and induction of T-regs. Extracellular matrix proteins formed regions within the LN that were permissive for co-localization of alloantigen-presenting cells, alloreactive T cells, and T-regs. Laminin-411 is produced by vascular endothelial and promotes T cell migration. Laminin-511 and laminin-521 are also present in the endothelial basement membrane. However, laminin-511 and fails to promote T cell migration, although T cell co-stimulatory properties have been reported. Here they identified unique expression patterns of laminin proteins that correlated with alloantigen-specific immunity or immune tolerance in mice. Laminin α4 (R&D Systems Inc.) and LN-511 were used for in vitro experiments. The ratio of laminin α4 to laminin α5 was greater in domains within tolerant LNs, compared with immune LNs, and blocking laminin α4 function or inducing laminin α5 overexpression disrupted T cell and dendritic cells localization These data again were commensurate with the in vitro migration results whereby laminin α5 impeded while α4 permitted migration through the endothelium and associated basement membrane. Furthermore, reducing α4 laminin circumvented tolerance induction and induced cardiac allograft inflammation and rejection in murine models. This work identifies laminins as potential targets for immune modulation.

  • 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.