Publications
Here is a selection of publications where different laminin isoforms were used to create more authentic cell culture systems.
In Vivo Generation of Post-infarct Human Cardiac Muscle by Laminin-Promoted Cardiovascular Progenitors
Yap L., Wang J.-W., Moreno-Moral A., de Kleijn D.P.V., Petretto E., Tryggvason K. Cell Reports, 2019
In this article, the authors report a high reproducible, chemically defined, xeno-free laminin-based differentiation protocol to generate stem cell-derived cardiovascular progenitors (CVPs). Laminin-221 (LN-221) was identified as the most likely expressed cardiac laminin. LN221 promoted the differentiation of pluripotent human embryonic stem cells (hESCs) toward cardiomyocyte lineage and downregulated pluripotency and teratoma-associated genes. Single-cell RNA sequencing of CVPs derived from hESC lines identified three main progenitor subpopulations. These CVPs were transplanted into myocardial infarction mice, where heart function was improved as measured by echocardiogram and human heart muscle bundle formation was identified histologically. This method may provide clinical-quality cells for use in regenerative cardiology.
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.
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-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.
CCAAT/enhancer binding protein-mediated regulation of TGFβ receptor 2 expression determines the hepatoblast fate decision
Takayama K., Kawabata K., Nagamoto Y., Inamura M., Ohashi K., Okuno H., Yamaguchi T., Tashiro K., Sakurai F., Hayakawa T., Okano T., Furue M.K., and Mizuguchi H. Development, 2014
Examined the function of TGFBR2 in the hepatoblast fate decision using hESC-derived HBC. hESC-derived HBCs purified and maintained (HBCs passaged more than three times) on human laminin 111 (LN111)-coated dishes were used. The HBC population was nearly homogeneous and expressed human hepatoblast markers such as alpha-fetoprotein (AFP), albumin (ALB), cytokeratin 19 (CK19) and EPCAM, and most of the colonies observed on human LN111-coated plates were ALB and CK19 double positive. The HBCs were capable of repopulating the liver of carbon tetrachloride (CCl4)-treated immunodeficient mice. This study reveals a molecular mechanism underlying the lineage commitment of human hepatoblasts (hepatocyte and biliary differentiation) controlled by a gradient of TGFβ signaling. It provides the first evidence of c/EBP-mediated regulation of TGFBR2 expression in the human hepatoblast fate decision.
Laminin-511 but not -332, -111, or -411 enables mouse embryonic stem cell self-renewal in vitro
Domogatskaya A., Rodin S., Boutaud A., Tryggvason K. Stem Cells, 2008
Different laminin isoforms, LN-511, -332, -411 and -111, as well as Matrigel, gelatin and poly-D-lysine are compared as substrates maintaining pluripotent mouse ES cells in vitro without the addition of leukemia inhibitory factor. Conclusions are that only LN-511 is able to sustain self-renewal for up to 169 days of culturing and cells maintain expression of pluripotency markers and can be used for generation of chimeric mice.
Integrin α6β1 Is the Main Receptor for Vascular Laminins and Plays a Role in Platelet Adhesion, Activation, and Arterial Thrombosis
Schaff et al.Circulation. 2013
Here, the authors show that laminin-411, laminin-511, and laminin-521, but not laminin-211, allow efficient platelet adhesion and activation across a wide range of arterial wall shear rates. Adhesion was critically dependent on integrin α6β1. Platelets from integrin α6 knockout mice failed to adhere to laminin-411, laminin-511, and laminin-521 but responded normally to a series of agonists. α6β1-Deficient mice presented a marked decrease in arterial thrombosis in 3 models of injury of the carotid, aorta, and mesenteric arterioles. The authors thus reveal an unsuspected important contribution of laminins to thrombus formation in vivo.
Contribution of α6 integrins to hematopoietic stem and progenitor cell homing to bone marrow and collaboration with α4 integrins
Qian H., Tryggvason K., Jacobsen S.E., Ekblom M.Blood, 2006
The integrin a6 chain is ubiquitously expressed in human and mouse hematopoietic stem and progenitor cells. The integrin a6 chain is ubiquitously expressed in human HPCs. Laminin-411 and -511, are present in subendothelial basement membranes of sinusoids in bone marrow, at sites of hematopoietic cell development and trafficking and might, therefore, regulate HSC functions. In this paper, they show that mouse HSC and progenitors express a6B1 integrin which mediates high cell adhesion to laminin-511 and 521 and to laminin-411 to a lower extent. Blocking of a6 significantly reduced progenitor cell homing to bone marrow in mice. Integrin a4 receptors are also important for homing of HSCs to bone marrow (but not to the spleen). The first data showing that a6 integrins (LN521/511 binding) function in vivo as hematopoietic stem and progenitor cell homing receptors. Also, show the role of integrin a4 receptor for homing of long-term multilineage reconstituting HSCs and collaboration of these 2 integrins in homing of short-term HSCs.