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

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  • Human Mesenchymal Cells from Adipose Tissue Deposit Laminin and Promote Regeneration of Injured Spinal Cord in Rats

    Menezes K., Nascimento M.A., Goncalves J.P., Silva Cruz A., Vieira Lopes D., Curzio B., Bonamino M., Ricardo J., de Menezes L., Borojevic R., Doria Rossi M.I., Coelho-Sampaio T. PLoS ONE, 2013

    In this article, regenerative properties of human adipose tissue derived stromal cells (hADSCs) were investigated in a rat model of spinal cord compression. hADSCs were shown to secrete laminin in the spinal cord. Out of six isoforms tested, they confirmed expression of α2 and α5 and also found positivity for β2 and γ1 laminin chains. The authors propose that laminin can be the paracrine factor mediating the pro-regenerative effects of hADSCs in spinal cord injury. This is supposedly the first article in which a link between laminin and the pro-regenerative effects of mesenchymal cells in the central nervous system is proposed.

  • Laminin α1 Chain Synthesis in the Mouse Developing Lung: Requirement for Epithelial–Mesenchymal Contact and Possible Role in Bronchial Smooth muscle Development

    Schuger L., Skubitz A.P.N., Zhang J., Sorokin L., He L. The Journal of Cell Biology, 1997

    The present study shows that whereas a1 and a2 laminins are synthesized in the mouse developing lung and in epithelial-mesenchymal co-cultures derived from it, epithelial and mesenchymal monocultures lose their ability to synthesize the laminin α1 chain. Synthesis of laminin α1 chain, however, returns upon re-establishment of epithelial-mesenchymal contact. Cell-cell contact is critical since laminin α1 chain is not detected in monocultures exposed to coculture-conditioned medium or in epithelial-mesenchymal cocultures in which heterotypic cell-cell contact is prevented by an interposing filter. Immunohistochemical studies on cocultures treated with brefeldin A, an inhibitor of protein secretion, indicated both epithelial and mesenchymal cells synthesize laminin α1 chain upon heterotypic cell-cell contact. Lung explants exposed to monoclonal antibodies to laminin α1 chain exhibited alterations in peribronchial cell shape and decreased smooth muscle development, as indicated by low levels of smooth muscle a actin and desmin. Taken together, the data suggest that laminin α1 chain synthesis is regulated by epithelial-mesenchymal interaction and may play a role in airway smooth muscle development.

  • Mesenchymal Stromal Cells for Sphincter Regeneration: Role of Laminin Isoforms upon Myogenic Differentiation

    Seeger T., Hart M., Patarroyo M., Rolauffs B., Aicher W.K., Klein G. PLoS One, 2015

    In the sphincter tissue (smooth muscle) a strong expression of the isoforms laminin-211/221, laminin-411/421 and laminin-511/521 can be detected in the different cell layers. Bone marrow-derived MSCs in culture, however, mainly express the isoforms laminin-411 and laminin-511, but not laminin-211. Even after myogenic differentiation, laminin-211 can hardly be detected. All laminin isoforms tested (-211, -411, -511 and -521) showed significant inhibition of the proliferation of undifferentiated MSCs but, with the exception of laminin-521, they had no influence on the proliferation of MSCs cultivated in a myogenic medium. The strongest cellular adhesion of MSCs was to laminin-511 and laminin-521, whereas laminin-211 was only a weakly-adhesive substrate for MSCs. Myogenic differentiation of MSCs even reduced the interaction with laminin-211, but it did not affect the interaction with laminin-511 and laminin-521. Since during normal myogenesis the latter two isoforms are the major laminins surrounding developing myogenic progenitors, α5 chain-containing laminins are recommended for further improvements of myogenic differentiation protocols of MSCs into smooth muscle cells.

  • Laminin-5 and type I collagen promote adhesion and osteogenic differentiation of animal serum-free expanded human mesenchymal stromal cells

    Mittag F., Falkenberg E-M, Janczyk A., Götze M., Felka T., Aicher W.K. Kluba T.Orthopedic Reviews, 2012

    In this article, the authors show that laminin 332 and type I collagen promote attachment and that laminin 332 promotes osteogenic differentiation of MSC. Expansion of MSC in animal serum-free, GMP-conforming media yielded vital cells meeting all minimal criteria for MSC. Attachment assay revealed a favorable binding of MSC to laminin 332 and type I collagen. Compared to plastic, osteogenic differentiation was significantly increased by laminin 332 after 28 days of culture, with no significant differences in gene expression patterns observed. Their data also confirm that laminin 332 serve better in bone repair, as this material promotes both firm attachment and osteogenic differentiation of MSC.

  • Regulation of Proliferation and Chondrogenic Differentiation of Human Mesenchymal Stem Cells by Laminin-5 (Laminin-332)

    Regulation of Proliferation and Chondrogenic Differentiation of Human Mesenchymal Stem Cells by Laminin-5 (Laminin-332)

    Hashimoto J., Hashimoto U., Kariya Y., Miyazaki K.
    STEM CELLS, 2006

    In this study, the authors examined a possible role of laminin 332 in the proliferation and differentiation of human MSCs. When MSCs were incubated in the presence of a coated or soluble form of laminin 332 in a growth medium, they proliferated more rapidly than nontreated cells, keeping their differentiation potential. On the other hand, laminin 332 potently suppressed the chondrogenic differentiation of MSCs. These activities were mediated mainly by integrin a3B1. However, laminin-5 had no effect on the osteogenic differentiation of MSCs. These results suggest that laminin-5 may contribute to the development of bone tissues by promoting proliferation and by suppressing the chondrogenic differentiation of MSCs.

  • Development of a biomaterial associated with mesenchymal stem cells and keratinocytes for use as a skin substitute

    Steffens D., Mathor M.B., Santi B.T.S., Luco D.P., Pranke P.Regen. Med., 2015

    Here they developed s scaffolds of poly-DL-lactic acid with and without the linkage of laminin-332, bringing together MSCs and keratinocytes aimed for treatment as a new skin substitute. Three groups of scaffolds were studied: 1) poly-DL-lactic acid (PDLLA), 2) hydrolyzed PDLLA (PDLLA/NaOH) and 3) PDLLA/Lam which is a PDLLA/NaOH scaffold linked to laminin-332. The results corroborate the hypothesis that laminin influenced the adhesion of the MSCs. Laminin significantly promoted the adhesion and spreading of proliferating oral and epidermal keratinocytes compared with collagen nanofibers only. The use of biocompatible and biodegradable polymers associated with the properties of laminin leads to an improvement in the adherence and viability of the cells, showing LN-332 is beneficial for the growth of MSCs and keratinocytes.

  • 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.

  • Laminin 411 acts as a potent inducer of umbilical cord mesenchymal stem cell differentiation into insulin-producing cells

    Qu H., Liu X., Ni Y., Jiang Y., Feng X., Xiao J., Guo Y., Kong D., Li A., Li X., Zhuang X., Wang Z., Wang Y., Chang Y., Chen S., Kong F., Zhang X., Zhao S., Sun Y., Xu D., Wang D., Zheng C. Journal of Translational Medicine, 2014

    Efficient induction of differentiation to insulin-producing cells from MSCs. Up-regulated insulin expression at both mRNA and protein levels. Administration of the insulin-producing cells in T1 diabetes rats rapidly 1) down-regulated fasting blood glucose levels, 2) significantly reduced the HbA1c concentration and 3) markedly improved the symptoms and survival of the rats.

  • CD49f Acts as an Inflammation Sensor to Regulate Differentiation, Adhesion, and Migration of Human Mesenchymal Stem Cells

    Yang Z., Dong P., Fu X., Li Q., Ma S., Wu D., Kang N., Liu X., Yan L., Xiao R.Stem Cells, 2015

    Here, we studied the role of CD49f (also known as integrin α6) in bone marrow MSCs. CD49f is preferentially expressed in fetal cells rather than adult cells, CD49f‐positive BM-MSCs possess higher CFU‐F formation ability and differentiation potential than CD49f negative cells, and the CD49f expression of BM-MSCs gradually decreases during in vitro passaging. An adhesion assay showed strong adhesion of BM-MSCs to both laminin 511 and 521 that were significantly higher than the control group coated with BSA, and the adhesion occurred evenly throughout the well. Pre‐blocking of CD49f on BM-MSCs inhibited the adhesion of fetal BM-MSCs to laminin 511 and 521. Also, CD49f knockdown dramatically decreased the differentiation of BMSCs. Inflammation (TNF-a) down-regulated CD49f in BMSCs with impaired differentiation, decreased adhesion to laminins and increased migration. This study provides evidence for CD49f as a stemness marker of BMSCs which is correlated with cell adhesion on laminin-521 and -511.

  • Bone Marrow Mesenchymal Stem Cells Adhesion Assay

    Yang Z. and Xiao R. Bio-protocol, 2016

    Here, the authors present a protocol for the culture of bone marrow MSC (BM-MSCs) on laminin-521 or laminin-511. The protocol is based on the method by Siler et al., 2000, and can easily be translated to MSCs from other origin or alternative ECMs coating. Both laminin isoforms show a significantly better efficient attachment compared to uncoated wells and also support seeding of a lower cell number compared to uncoated plats. The BM-MSCs adhere to the laminin-coated wells within 10 min, while for non-coated wells, it may take a longer time. In this protocol, a final coating concentration of 2 μg/cm2 is used but can effectively be lowered 4-10 times without loss of function.