Publications

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

  • Area of interest

  • Transcriptomes of germinal zones of human and mouse fetal neocortex suggest a role of extracellular matrix in progenitor self-renewal

    Fietz et al.
    PNAS. 2012.

  • Laminin enhances the growth of human neural stem cells in defined culture media

     

    Hall et al.
    BMC Neuroscience, 2008

  • Adult SVZ Stem Cells Lie in a Vascular Niche: A Quantitative Analysis of Niche Cell-Cell Interactions

    Shen Q., Wang Y., Kokovay E., Lin G., Chuang S-M. Goderie S.K., Roysam B., Temple S. Cell Stem Cell, 2008

    Here, they examine the relationship of adult SVZ NSC lineage cells to blood vessels using confocal imaging of SVZ whole mounts in which the normal 3D relationships of cells are preserved. Neural stem cells (NSCs) in the adult subventricular zone (SVZ) lie close to blood vessels in a vascular-derived laminin-rich niche. Cells expressing stem cell markers, including GFAP, and proliferation markers are closely apposed to the laminin-containing extracellular matrix (ECM) surrounding vascular endothelial cells. Adult SVZ progenitor cells express the laminin receptor a6B1 integrin, and blocking this inhibits their adhesion to endothelial cells, altering their position and proliferation in vivo.

  • Laminin and growth factor receptor activation stimulates differential growth responses in subpopulations of adult DRG neurons

    Tucker B.A., Rahimtula M., Mearow K.M.European Journal of Neuroscience, 2006

    Here they show laminin-induced neurite outgrowth and its relation to three known DRG neuronal types. They also show PI3K pathway is responsible. They also discuss this in the light of possible therapeutic targets. The study is limited in that that they only use invitrogen laminin (purified laminin-111) and isoform-specific effects cannot be seen, but they come to a number of highly interesting conclusions: 1) The current findings provide strong support for the use of the ECM molecule laminin in conjunction with NGF and GDNF in order to stimulate optimal levels of axon growth from all populations of regenerating sensory neurons. 2) identified intracellular signaling components that provide potential therapeutic targets when attempting to stimulate the regeneration of peripheral axons. Pharmacological alterations of the PI 3-K/Akt pathway resulting in activation of either PI 3-K or Akt could be beneficial. 3) Laminin-induced neurite growth occurs in the absence of added trophic factors only in heavy-chain neurofilament-positive and calcitonin gene-related peptide-positive DRG neurons [nerve growth factor (NGF)-responsive population]. In contrast, laminin alone is not sufficient to stimulate significant neurite growth from lectin Griffonia simplicifolia IB4-positive neurons (IB4+ve), although it is still required to elicit a growth response from these cells in the presence of glial-derived neurotrophic factor. 

  • Quantification of molecular interactions between ApoE, amyloid-beta (Aβ) and laminin: Relevance to accumulation of Aβ in Alzheimer's disease

    Zekonyte J., Sakai K., Nicoll J.A.R., Weller R.O., Carare R.O.Biochimica et Biophysica Acta, 2015

    Accumulation of amyloid-β (Aβ) in plaques in the brain and in artery walls as cerebral amyloid angiopathy indicates a failure of elimination of Aβ from the brain with age and Alzheimer's disease. A major pathway for elimination of Aβ and other soluble metabolites from the brain is along basement membranes within the walls of cerebral arteries that represent the lymphatic drainage pathways for the brain. Since Aβ40 is the predominant type of Aβ found in cerebral amyloid angiopathy, in the present study we tested the hypothesis that interactions of Aβ40 with protein components of cerebral vascular basement membranes, such as laminin, are stronger in the presence of ApoE3 than in the presence of ApoE4. Proteins (LN-511, ApoE3, or ApoE4) were immobilized on AFM probes using the amine–amine-reactive linker aldehyde–PEG–NHS. Force-spectroscopy experiments were performed to study the reciprocal influence of different isoforms of ApoE and Aβ on their binding interactions with laminin-511. The results show that apolipoprotein E co-localizes with Aβ in basement membrane drainage pathways in the walls of arteries. Moreover, all AFM measurements demonstrate that Aβ + ApoE3 complex has a stronger binding to laminin-511 than Aβ + ApoE4. These results suggest that perivascular elimination of ApoE4/Aβ complexes would be less efficient than with other isoforms of apolipoprotein E, thus endowing a higher risk for Alzheimer's disease. Therapeutic correction for ApoE4/Aβ/laminin interactions may increase the efficiency of the elimination of Aβ in the prevention of Alzheimer's disease.

  • Laminin-411 Is a Vascular Ligand for MCAM and Facilitates TH17 Cell Entry into the CNS

    Flanagan K., Fitzgerald K., Baker J., Regnstrom K., Gardai S., Bard F., Mocci S., Seto P., You M., Larochelle C., Prat A., Chow S., Li L., Vandevert C., Zago W., Lorenzana C., Nishioka C., Hoffman J., Botelho R., Willits C., Tanaka K., Johnston J., Yednock T.PLOS ONE, 2012

    TH17 cells enter tissues to facilitate pathogenic autoimmune responses. Herein, they characterize MCAM (CD146) as an adhesion molecule that defines human TH17 cells. Parental CHO cells, lacking MCAM expression or CHO cells stably transfected with mouse MCAM were incubated in the presence of laminin-411 or laminin-511. They identify the MCAM ligand as laminin 411, an isoform of laminin expressed within the vascular endothelial basement membranes. hMCAM binds to a ligand in the ECM with identical staining to laminin a4. Moreover, mMCAM colocalizes with laminin 411 on the choroid plexus, and shows no specific binding to tissues from LAMA4 -/- mice. Their data suggest that MCAM and laminin-411 interact to facilitate TH17 cell entry into tissues and promote inflammation. The specific location of laminin 411 in the endothelial basement membrane may either function to augment adhesion of cells attempting CNS endothelial penetration or serve as an adhesion based gating system to signal appropriate entry mechanisms. As such, modulation of the interaction between MCAM and laminin 411 represents a novel and selective approach that may help to maintain or restore homeostasis to inflamed tissues in autoimmune diseases.

  • Robust Formation and Maintenance of Continuous Stratified Cortical Neuroepithelium by Laminin-Containing Matrix in Mouse ES Cell Culture

    Nasu M., Takata N., Danjo T., Sakaguchi H., Kadoshima T., Futaki S., Sekiguchi K., Eiraku M., Sasai Y. PLoS ONE, 2012

    Here the authors report substantial supporting effects of the extracellular matrix (ECM) protein laminin on the continuous formation of properly polarized cortical NE in a floating aggregate culture of mESCs. The laminin protein used here contained laminin 111 that was generated from cultured cells and co-purified with entactin (nidogen-1/2), a laminin-binding protein. The addition of purified laminin and entactin, even at low concentrations, stabilized the formation of continuous cortical NE as well as the maintenance of the basement membrane and prevented rosette formation. The results indicate that laminin-entactin ECM promotes the formation of structurally stable telencephalic tissues in 3D mESC culture, and supports the morphogenetic recapitulation of cortical development.

  • Molecular Diversity of Midbrain Development in Mouse, Human, and Stem Cells

    Manno, Gyllborg, Codeluppi, Nishimura, Salto, Zeisel, Borm, Stott, Toledo, Villaescusa, Lönnerberg, Ryge, Barker, Arenas, Linnarsson.Cell, 2016

    Manno and colleagues used single-cell RNA sequencing to examine ventral midbrain development in humans and mice. They found that cell types and gene expression were generally conserved across species, but with clear differences in cell proliferation, developmental timing, and dopaminergic neuron development. Additionally, they quantitatively assessed the fidelity of dopaminergic neurons derived from human pluripotent stem cells, at a single-cell level. The study provides insight into the molecular programs controlling human midbrain development and provides a foundation for the development of cell replacement therapies.

  • Laminin targeting of a peripheral nerve-highlighting peptide enables degenerated nerve visualization

    Glasgow H.L., Whitney M.A., Gross L.A., Friedman B., Adams S.R., Crisp J.L., Hussain T., Frei A.P., Novy K., Wollscheid B., Nguyen Q.T., Tsien R.Y.PNAS, 2106

    Here, the extracellular matrix proteins laminin-421 and -211 were identified as NP41 binding targets, and TRICEPS-based glycoprotein capture supported laminin-421 as the primary binding target. Fluorescently labeled nerve-binding peptide NP41 holds promise to reduce surgical nerve damage and facilitate nerve repair. Clinical translation hinges on the identification of binding targets to assess potential toxicity and understand the mechanism. For target identification, the authors developed a receptor capture method, enabling covalent tagging and identification of proteins within close proximity to a bound ligand. The results explain the ability of NP41 to highlight degenerated nerve “ghosts” months after transection that were invisible to the unaided eye but contain laminins. Targeting the extracellular matrix is advantageous for clinical imaging agents, likely reducing undesirable neurological effects. 

  • Human mesenchymal cells from adipose tissue deposit laminin and promote regeneration of injured spinal cord in rats

    Menezes K., Nascimento M.A., Gonçalves J.P., Cruz A.S., Lopes D.V., Curzio B., Bonamino M., de Menezes J.R., Borojevic R., Rossi M.I., Coelho-Sampaio T.PLOS ONE, 2014

    Here the authors investigated the regenerative properties of human adipose tissue-derived stromal cells (hADSCs) in a rat model of spinal cord compression. Cells were delivered directly into the spinal parenchyma immediately after the injury. Human ADSCs promoted functional recovery, tissue preservation, and axonal regeneration. Analysis of the cord tissue showed an abundant deposition of laminin of human origin at the lesion site and spinal midline; the appearance of cell clusters composed of neural precursors in the areas of laminin deposition, and the appearance of blood vessels with separated basement membranes along the spinal axis. These effects were also observed after the injection of hADSCs into the non-injured spinal cord. Considering that laminin is a well-known inducer of axonal growth, as well a component of the extracellular matrix associated with neural progenitors, the authors propose that it can be the paracrine factor mediating the pro-regenerative effects of hADSCs in spinal cord injury.