Publications

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

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  • Leveraging interacting signaling pathways to robustly improve the quality and yield of human pluripotent stem cell-derived hepatoblasts and hepatocytes

    Claudia Raggi, Marie-Agnès M’Callum, Quang Toan Pham, Perrine Gaub, Silvia Selleri, Nissan Vida Baratang, Chenicka Lyn Mangahas, Gaël Cagnone, Bruno Reversade, Jean-Sébastien Joyal, Massimiliano Paganelli. Stem Cell Reports, 2022

    This study describes a robust and scalable protocol for pluripotent stem cell (PSC) differentiation into high-quality bipotent hepatoblasts and hepatocyte-like cells (HLCs), with Biolaminin 521 (laminin-521) as the cell culture matrix. The protocol resulted in cells that had a better resemblance to primary human hepatocytes than protocols described previously, showing a good potential to use them as an alternative to primary cells for in vitro modeling. PSC-derived hepatoblasts were able to mature into hepatocytes and bile ducts within syngeneic liver organoids.

  • Impaired integrin α5β1-mediated hepatocyte growth factor release by stellate cells of the aged liver

    Rohn F., Kordes C., Buschmann T., Reichert D., Wammers M., Poschmann G., Stühler K., Benk A.S., Geiger F., Spatz J.P., Häussinger D.Aging Cell, 2020

    Laminin proteins are critically involved in HSC function which is further illustrated in this article. Here, the authors illustrate the mechanistic link between fluid mechanical forces, loss of important extracellular matrix proteins, such as laminins, and hepatic aging, and how it all leads to an impaired liver regeneration potential. The authors provide evidence that integrin α5β1 is an important mechanosensor in hepatic stellate cells (HSC) involved in shear stress-induced release of hepatocyte growth factor (HGF), an essential inductor of liver regeneration which is impaired during aging. The expression of the integrin subunits α5 and β1 decreases in liver and HSC from aged rats. CRISPR/Cas9-mediated integrin α5 and β1 knockouts in isolated HSC lead to lowered HGF release and impaired cellular adhesion. Fluid mechanical forces increase integrin α5 and laminin gene expression whereas integrin β1 remains unaffected. In the aged liver, laminin β2 and γ1 protein chains as components of laminin-521 are lowered. The integrin α5 knockout in HSC reduces laminin expression via mechanosensory mechanisms. Culture of HSC on nanostructured surfaces functionalized with laminin-521 enhances HGF expression in HSC, demonstrating that these laminin proteins are critically involved in HSC function. During aging, HSC acquires a senescence-associated secretory phenotype and lower their growth factor expression essential for tissue repair. These findings suggest that impaired mechanosensing via integrin α5β1 in HSC contributes to the age-related reduction of ECM and HGF release that could affect liver regeneration.

  • Recombinant Laminins Drive the Differentiation and Self-Organization of hESC-Derived Hepatocytes

    Cameron K., Tan R., Schmidt-Heck W., Campos G., Lyall M.J, Wang Y., Lucendo-Villarin B., Szkolnicka D., Bates N., Kimber S.J., Hengstler J.G., Godoy P., Forbes S.J., Hay D.C. Stem Cell Reports, 2015

    Human ES cells are cultured on human recombinant laminin-521 and laminin-111 shows efficient hepatocyte specification, maturation, function and stabilization of phenotype. The results presented in the paper represent a significant advance compared to any previously published data especially regarding metabolic activity and functional organization. Cells cultured on the laminin matrices exhibited significantly increased metabolic function relative to cells on Matrigel and human primary hepatocytes. The laminin cultured hepatocyte-like cells were arranged in lobule like structures, reminiscent of regenerating liver, with positive staining for MRP1 and MRP2 and were capable of biliary efflux.

  • Protocol: Defined and Scalable Generation of Hepatocyte-like Cells from Human Pluripotent Stem Cells

    Wang Y., Alhaque S., Cameron K., Meseguer-Ripolles J., Lucendo-Villarin B., Rashidi H., Hay D.C. JoVE, 2017

    Here, the authors show in a video how they developed a defined optimized system for hepatocyte differentiation using human recombinant laminins as extracellular matrices in combination with a serum-free differentiation process. The protocol shows the procedures for culturing hPSCs on LN521 and differentiating them on either LN521 or a blend of LN521 and LN111 (Biolaminin 521 and 111). Highly efficient hepatocyte specification was achieved, with demonstrated improvements in both HLC function and phenotype. Importantly, this system is easy to scale up using research and GMP-grade hPSC lines promising advances in cell-based modeling and therapies.

  • Long-Term Self-Renewal of Human ES/iPS-Derived Hepatoblast-like Cells on Human Laminin 111-Coated Dishes

    Takayama K., Nagamoto Y., Mimura N., Tashiro K., Sakurai F., Tachibana M., Hayakawa T., Kawabata K., Mizuguchi H.  Cell Stem Cell Reports, 2013 

    The authors of this important study demonstrated that laminin-111 is the optimal matrix for hepatoblasts. In addition to solving the need to efficiently expanding and purifying this liver progenitor, the matrix provides an important safety mechanism as LN-111 selectively only maintains hepatoblasts while eliminating residual undifferentiated cells that cannot survive and self-renew on laminin-111.

  • Maintenance of Hepatic Functions in Primary Human Hepatocytes Cultured on Xeno-Free and Chemical Defined Human Recombinant Laminins

    Watanabe M., Zemack H., Johansson H., Hagbard L., Jorns C., Li M., Ellis E.PLOS ONE, 2016

    In this study, the authors determined the distribution of laminin isoforms in human liver tissue and isolated primary human hepatocytes and investigated the efficacy of different human recombinant laminin isoforms on hepatic functions during culture. Protein expressions of laminin-chain α2, α3, α4, β1, β3, γ1, and γ2 were detected in both isolated human hepatocytes and liver tissue. α1 and α5 expression could be detected on RNA level but not on protein level in liver tissue or hepatocytes. Hepatocytes were isolated from five different individual livers, and cultured on human recombinant laminin isoforms -111, -211, -221, -332, -411, -421, -511, and -521, matrigel or collagen type IV. Hepatocytes cultured on laminin showed characteristic hexagonal shape in a flat cell monolayer. Viability, double-stranded DNA concentration, and Ki67 expression for hepatocytes cultured for six days on laminin were comparable to those cultured on EHS and Collagen. Hepatocytes cultured on laminin also displayed production of human albumin, alpha-1-antitrypsin, bile acids, and gene expression of liver-enriched factors, such as hepatocyte nuclear factor 4 alpha, glucose-6-phosphate, cytochrome P450 3A4, and multidrug resistance-associated protein 2. We conclude that all forms of human recombinant laminin tested maintain cell viability and liver-specific functions of primary human hepatocytes and that recombinant laminin is a promising xeno-free and chemical defined strategy for preservation of hepatocyte-specific function in vitro.

  • 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.

  • Prediction of interindividual differences in hepatic functions and drug sensitivity by using human iPS-derived hepatocytesCell Stem Cell Reports, 2013 Oct

    Takayama K., Morisaki Y., Kuno S., Nagamoto Y., Harada K., Furukawa N., Ohtaka M., Nishimura K., Imagawa K., Sakurai F., Tachibana M., Sumazaki R., Noguchi E., Nakanishi M., Hirata K., Kawabata K., Mizuguchi H.  PNAS 2014

    The authors had previously developed a method to maintain and proliferate PSC-derived hepatoblasts on LN-111 (Cell Stem Cell Reports, 2013 Oct). In this publication, they examine and find evidence for the increased efficiency and homogeneity of hepatocyte differentiation when the LN-111 cultivated and purified hepatoblasts are further differentiated on LN-111 to hepatocytes.

  • A Chemically Defined Hydrogel for Human Liver Organoid Culture

    Ye S., Boeter J.W.B., Mihajlovic M., van Steenbeek F.G, van Wolferen M.E., Oosterhoff  L.A., Marsee A., Caiazzo M., van der Laan L.J.W., Penning L.C., Vermonden T., Spee B., and Schneeberger K. Adv. Funct. Mater. 2020

    Here, a novel hydrogel-based on polyisocyanopeptides (PIC) and Biolaminin-111 is described for human liver organoid cultures. PIC is a synthetic polymer that can form a hydrogel with thermosensitive properties, making it easy to handle and very attractive for clinical applications. PIC hydrogel alone was not sufficient to support organoid growth. The addition of a laminin-entactin complex (LEC) to the plain PIC gel, resulted in efficient organoid formation and proliferation that seemed comparable to the Matrigel controls, with lower stiffnesses most favorable for organoid proliferation. The stem cell phenotype and proliferation and differentiation capacity of the organoids could be maintained in PIC-LEC over several passages, enabling their seemingly unlimited expansion and subsequent maturation. Moreover, organoids can be efficiently differentiated toward a hepatocyte-like phenotype with key liver functions. Importantly, they also show that the LEC in the PIC-LEC gels could be replaced by Biolaminin-111, resulting in a completely synthetic hydrogel for the expansion of human liver organoids.

  • Laminin and b1 Integrins Are Crucial for Normal Mammary Gland Development in the Mouse

    Klinowska T.C.M., Soriano J.V., Edwards G.M., Oliver J.M., Valentijn A.J., Montesano R., Streuli C.H.Developmental Biology, 1999

    Here, the authors examine the role of integrin-extracellular matrix interactions in the morphogenesis of ductal structures in vivo (mouse). End buds are surrounded by a basement membrane, which is shown to contain laminin-111 and collagen IV. Blocking B1 integrins dramatically reduced both the number of end buds per gland and the extent of the mammary ductal network, compared with controls. These effects were specific to the end buds since the rest of the gland architecture remained intact. Similar results were obtained with anti-laminin antibodies. In contrast, no effect on morphogenesis in vivo was seen with anti-a6 integrin antibody, suggesting that a6 is not the important partner for b1 in this system. They also show that integrins and hepatocyte growth factor (HGF) cooperate to regulate ductal morphogenesis. We propose that both laminin and b1 integrins are required to permit cellular traction through the stromal matrix and are therefore essential for maintaining end bud structure and function in normal pubertal mammary gland development.