Publications
Here is a selection of publications where different laminin isoforms were used to create more authentic cell culture systems.
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.
Aligned Poly(ε-caprolactone) Nanofibers Guide the Orientation and Migration of Human Pluripotent Stem Cell-Derived Neurons, Astrocytes, and Oligodendrocyte Precursor Cells In Vitro
Hyysalo A., Ristola M., Jpki T., Honkanen M., Vippola M., Narkilahti S.Macromolecular Bioscience, 2017
Biomaterials can be used as supporting scaffolds, as they mimic the characteristics of the natural cellular environment. In this study, hPSC-derived neurons, astrocytes, and oligodendrocyte precursor cells (OPCs) are cultured on aligned poly(ε-caprolactone) nanofiber platforms, which guide cell orientation to resemble that of the spinal cord in vivo. Human neurons and astrocytes require extracellular matrix molecule coating for the nanofibers, but OPCs grow on nanofibers without additional treatment. Clinically relevant human recombinant laminin substrates (laminin-521 and laminin-511) are compatible with PCL nanofibers and can be used for efficient culturing of hPSC-derived neurons and astrocytes on PCL nanofibers. Furthermore, the nanofiber platform is combined with a 3D hydrogel scaffold with controlled thickness, and the nanofiber-mediated orientation of hPSC-derived neurons is also demonstrated in a 3D environment.
Laminin α5 substrates promote survival, network formation and functional development of human pluripotent stem cell-derived neurons in vitro
Anu Hyysalo, Mervi Ristola, Meeri E.-L. Mäkinen, Sergei Häyrynen, Matti Nykter, Susanna Narkilahti. Stem Cell Research, 2017
In this work, the authors compared different recombinant human laminin isoforms in the culture of hPSC derived neurons. They show that LN α5 chain is important for neuronal attachment, viability and network formation, and that neuronal functional development is enhanced by LN α5-substrates.
Laminin 521 maintains differentiation potential of mouse and human satellite cell-derived myoblasts during long-term culture expansion
Penton C.M., Badarinarayana V., Prisco J., Powers E., Pincus M., Allen R.E., August P.R. Skeletal Muscle, 2016
Here, the authors comprehensively examine the effect of physiologically relevant laminins, laminin-211 and laminin-521, compared to traditionally utilized ECMs (e.g., laminin-111, gelatin, and Matrigel) to assess their capacity to propagate and preserve myogenic differentiation potential. The results demonstrate laminin-521 is a superior substrate for both short-term and long-term myogenic cell culture applications compared to other commonly utilized substrates. Laminin-521 also provides more consistent and reliable differentiation over long-term culture. Laminin-521 supported increased proliferation in early phases of expansion and was the only substrate facilitating high-level fusion following eight passages in mouse myoblast cell cultures. In human myoblast cell cultures, laminin 521 supported increased proliferation during expansion and superior differentiation with myotube hypertrophy. Counterintuitively, however, laminin-211, the native laminin isoform in resting skeletal muscle, resulted in the low proliferation and poor differentiation in mouse and human cultures. Matrigel performed well in short-term mouse studies but showed high amounts of variability following long-term expansion.
iPSC-derived human cardiac progenitor cells improve ventricular remodelling via angiogenesis and interstitial networking of infarcted myocardium
Ja K.P., Miao Q., Zhen Tee N.G., Lim S.Y., Nandihalli M, Ramachandra C.J.A, Mehta A, Shim W. J Cell Mol Med. 2015
Here they investigate the effects of myocardial transplantation of human induced pluripotent stem cell (iPSC)-derived progenitors and cardiomyocytes into acutely infarcted myocardium in immune deficient mice. iPSC cultured on Matrigel and differentiated in EB structures. Differences in integrin and laminin expression between cardiac progenitors and cardiomyocytes were observed. The a6 integrin was higher expressed in progenitors and integrin a1, a2, a3, a7 and b1 higher expression in cardiomyocytes. They observed a distinct swish in expression profile of laminin subunits during cardiac differentiation with laminin-411/421 pre-dominantly expressed early in progenitors and laminin-211/221 expressed later in cardiomyocytes. Improvements of myocardial function in the progenitor group corresponded to increased vascularization and coincided with augmented networking of cardiac telocytes in the interstitial space of the infarcted zone. Laminin-221/211-expressing cardiomyocytes only retained and engrafted around myofibres in the peri-infarct region.