Publications
Here is a selection of publications where different laminin isoforms were used to create more authentic cell culture systems.
Publication: Single-cell transcriptomics captures features of human midbrain development and dopamine neuron diversity in brain organoids
Fiorenzano et al. 2021 Nature Communications
Biosilk 3D scaffold biofunctionalized with Biolaminin 111 was used to generate mature and functional ventral midbrain (VM) organoids with less variation both between the organoid replicates and within an organoid, when compared with a standard VM-organoid method. This study shows the benefits of Biosilk especially in terms of permeability (no necrotic centers), resulting in improved cell maturity and experimental reproducibility. Biosilk can be supplemented with any Biolaminin isoforms, depending on the tissue type of interest.
A Hydrogel Platform that Incorporates Laminin Isoforms for Efficient Presentation of Growth Factors – Neural Growth and Osteogenesis
Oana Dobre, Mariana A. G. Oliva, Giuseppe Ciccone, Sara Trujillo, Aleixandre Rodrigo-Navarro, Douglas Cormac Venters, Virginia Llopis-Hernandez, Massimo Vassalli, Cristina Gonzalez-Garcia, Matthew J. Dalby, Manuel Salmeron-Sanchez. Advanced Functional Materials, 2021
The authors report a 3D culture system with a defined matrix composition that reflects the complexity of the native ECM, where growth factors in combination with Biolaminin isoforms give more natural cellular processes. The authors incorporated the full-length Biolaminin 521, 332, and 411 proteins into a synthetic polymer network with controlled physico-chemical properties, and showed examples of hMSC osteogenesis and neurite growth in this 3D microenvironment.
Protocol for automated production of human stem cell derived liver spheres
Jose Meseguer-Ripolles, Alvile Kasarinaite, Baltasar Lucendo-Villarin, David C Hay. STAR Protoc, 2021
In this article, the authors describe how they produce human liver spheres from pluripotent stem cell-derived hepatic progenitors, endothelial cells, and hepatic stellate cells, using LN521 in the differentiation protocol. Their process is automated using liquid handling and pipetting systems, permitting cost-effective scale-up and reducing sphere variability.
Publication: Biosilk with laminin-521 for hPSC neural differentiation in 3D network
Åstrand et al. 2020 Biomaterials Science
This article describes the use of a recombinant spider silk protein functionalized with a cell binding motif from fibronectin in combination with a human recombinant laminin 521 (LN-521) to create a fully defined stem cell niche in 3D. The results show that hPSCs integrated into the foam develop into neural progenitors and that they stay viable during long-term differentiations. The culture system also supports morphogenesis mimicking the human brain development and can serve as base for engineering of hPSC-derived neural tissue. The article describes the 3D culture matrix sold under the name Biosilk®.
Publication: Assembly of functionalized silk together with cells to obtain proliferative 3D cultures
Johansson et al. 2019 Scientific Reports
This article describes the use of fibrous spider silk 3D network (Biosilk) for the culture of mammalian cells. The cells get uniformly integrated between the formed microfibers and the cells are highly proliferative, spreading out more efficiently than when encapsulated in a hydrogel. The authors conclude that the silk-assembly in presence of cells constitutes a viable option for 3D culture of cells integrated in an ECM-like network.
Laminin as a Potent Substrate for Large-Scale Expansion of Human Induced Pluripotent Stem Cells in a Closed Cell Expansion System
Gjorevski N. & Lutoff MP. Nat Protoc, 2017
In this work, the authors describe a protocol for the generation of well-defined matrices for the culture of intestinal stem cells (ISCs) and intestinal organoids, using PEG hydrogel backbone functionalized with minimal adhesion cues including RGD (Arg-Gly-Asp), which is sufficient for ISC expansion, and laminin-111, which is required for organoid formation.
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.
Collaboration of 3D Context and Extracellular Matrix in the Development of Glioma Stemness in a 3D Model
Ma N.K.L., Kai Lim J., Fatt Leong M., Sandanaraj E., Ti Ang B., Tang C., Wan A.C.A. Biomaterials, 2015
This work illustrates that different laminin isoforms have specific effects in promoting the stemness of glioma cells, in collaboration with a 3D context. U251 glioblastoma cells were cultured on electrospun polystyrene (ESPS) scaffolds coated with 7 different laminin isoforms (LAMscreen kit) to provide a 3D model for stem cell-related genes and proteins expression studies. The results indicate the influence of 3D (versus 2D) context on stemness markers and integrin expression, specifically, the upregulation of the laminin-binding integrins a6b4. By a colony-forming assay, we showed enhanced clonogenicity of cells grown on ESPS scaffolds in collaboration with laminins 411, 421, 511 and 521. The present results demonstrate how 3D versus 2D context profoundly affects ECM signaling, leading to stemness.
Designer matrices for intestinal stem cell and organoid culture
Gjorevski N., Sachs N., Manfrin A., Giger S., Bragina M.E., Ordóñez-Morán P., Clevers H., Lutolf M.P. Nature letters, 2016
Here the authors used modular synthetic hydrogel (cross-linked poly(ethylene glycol) (PEG)) to define the key extracellular matrix (ECM) parameters that govern intestinal stem cell (ISC) expansion and organoid formation, and show that separate stages of the process require different mechanical environments and ECM components. Fibronectin-based adhesion was sufficient for ISC survival and proliferation and high matrix stiffness significantly enhanced ISC expansion through a yes-associated protein 1 (YAP)-dependent mechanism. ISC differentiation and organoid formation, on the other hand, required a soft matrix and full-length laminin-111-based adhesion. The authors also produced mechanically dynamic matrices that were initially optimal for ISC expansion and subsequently permissive to differentiation and intestinal organoid formation.
Gelatine methacrylamide-based hydrogels – an alternative 3D cancer cell culture system
Kaemmerer E., Melchels F.P.W, Holzapfel B.M, Meckel T., Hutmacher D.W., Loessner D. Acta Biomaterialia, 2014
The authors present a 3D biomaterial platform for the analysis of ovarian cancer spheroid growth that is an efficient semi-synthetic alternative, combining native ECM components and tunable matrix properties, resulting in higher reproducibility, less complexity and better comparability between different groups than traditional cell monolayer approaches. In this study, gelatine methacrylamide-based hydrogels (GelMA) with added LN-411 were established as in vitro and in vivo spheroid-based 3D cancer models.
