Publications
Here is a selection of publications where different laminin isoforms were used to create more authentic cell culture systems.
Laminin enhances the growth of human neural stem cells in defined culture media
Hall et al.
BMC Neuroscience, 2008Protocol for the derivation, culturing, and differentiation of human iPS-cell-derived neuroepithelial stem cells to study neural differentiation in vitro
Javier Calvo-Garrido, Dania Winn, Camilla Maffezzini, Anna Wedell, Christoph Freyer, Anna Falk, Anna Wredenberg. STAR Protocols, 2021
This protocol describes the derivation of neuroepithelial stem (NES) cells from human induced pluripotent stem cells. NES cells can be further differentiated into neurons and glia. The PSC culture and NES differentiation were done on plates coated with Biolaminin 521 (laminin-521). To avoid clonal selection of isolated NES cells, it is recommended to follow the culture conditions described. The protocol is expected to result in highly proliferate NES cells providing a good source of cells of a neuronal cell lineage. Glial cells are formed after approximately 45 days of differentiation.
Human iPS-Derived Astroglia from a Stable Neural Precursor State Show Improved Functionality Compared with Conventional Astrocytic Models
Anders Lundin, Louise Delsing, Maryam Clausen, Piero Ricchiuto, José Sanchez, Alan Sabirsh, Mei Ding, Jane Synnergren, Henrik Zetterberg, Gabriella Brolén, Ryan Hicks, Anna Herland, and Anna Falk Stem Cell Reports, 2018
This study reports the differentiation of human induced pluripotent stem cell (hiPSC)-derived astroglia cells under defined conditions using Biolaminin 521 as the matrix.
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.
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®.
Direct Reprogramming of Human Fetal- and Stem Cell-Derived Glial Progenitor Cells into Midbrain Dopaminergic Neurons
Nolbrant S., Giacomoni J., Hoban D.B, Bruzelius A., Birtele M., Chandler-Militello D., Pereira M., Rylander Ottosson D., Goldman S.A., Parmar M. Stem Cell Reports, 2020
Human glial progenitor cells (hGPCs) are promising cellular substrates to explore for the in situ production of new neurons for brain repair. Proof of concept for direct neuronal reprogramming of glial progenitors using human cells has been difficult to perform since hGPCs are born late during human fetal development, with limited accessibility for in vitro culture. In this study, the authors provide evidence that hGPCs isolated from both the human fetal brain and differentiated from hESCs can be successfully reprogrammed into functional iNs, including induced DA neurons (iDANs). They also establish a renewable and reproducible stem cell-based hGPC system for direct neural conversion in vitro. Using this system, they have identified optimal combinations of fate determinants for the efficient dopaminergic (DA) conversion of hGPCs, thereby yielding a therapeutically relevant cell type that selectively degenerates in Parkinson’s disease.
Functional characterization of human pluripotent stem cell-derived cortical networks differentiated on laminin-521 substrate: comparison to rat cortical cultures
Tanja Hyvärinen, Anu Hyysalo, Fikret Emre Kapucu, Laura Aarnos, Andrey Vinogradov, Stephen J Eglen, Laura Ylä-Outinen, Susanna Narkilahti. Sci Rep, 2019
In this article, differentiation of functionally active hPSC-derived cortical networks on defined laminin-521 substrate is reported. They assessed compared the activity development of hPSC-derived networks to that of widely used rat embryonic cortical cultures using microelectrode array (MEA) measurements. The authors conclude that hPSC-derived neural cultures produced with a defined protocol generate cortical type network activity, and they could be applied as a human-specific model for pharmacological studies and modeling network dysfunctions.
Efficiently Specified Ventral Midbrain Dopamine Neurons from Human Pluripotent Stem Cells Under Xeno-Free Conditions Restore Motor Deficits in Parkinsonian Rodents
Jonathan C Niclis, Carlos W Gantner, Walaa F Alsanie, Stuart J McDougall, Chris R Bye, Andrew G Elefanty, Edouard G Stanley, John M Haynes, Colin W Pouton, Lachlan H Thompson, Clare L Parish. Stem Cells Transl Med, 2017
The authors describe the first fully defined feeder- and xenogeneic-free protocol for the generation of vmDA neurons from hPSCs for use in animal models of Parkinson's disease. This protocol, utilizing Biolaminin 521 as a component, consistently increases both the yield and proportion of vmDA neural progenitors and neurons that display classical vmDA metabolic and electrophysiological properties. These findings may help in translation of hPSC-derived neurons into the clinic.
Neurons From Human Pluripotent Stem Cells Under Xeno-Free Conditions Restore Motor Deficits in Parkinsonian Rodents
Niclis J.C., Gantner C.W., Alsanie W.F., McDougall S.J., Bye C.R., Elefanty A.G., Stanley E.G., Haynes J.M., Pouton C.W., Thompson L.H., Parish C.L.Stem cells translational medicine, 2016
In this study, the authors describe the first fully defined feeder- and xenogeneic-free protocol for the generation of vmDA neurons from hPSCs. The protocol is translational across multiple embryonic and induced hPSC lines. hPSCs were cultured xeno-free on laminin-521 in TeSR2. For vmDA differentiation, two xeno-free matrix proteins, vitronectin, and human laminin-521 were compared for their ability to replace Matrigel. Both matrices facilitated appropriate patterning, however, only laminin-521 supported the long-term attachment of neural precursors. This “next-generation” protocol consistently increases both the yield and proportion of vmDAneural progenitors (OTX2/FOXA2/LMX1A) and neurons (FOXA2/TH/PITX3) that display classical vmDA metabolic and electrophysiological properties. The mechanism underlying these improvements are identified and demonstrate clinical applicability with the first report of scalability and cryopreservation of bona fide vmDA progenitors at a time amenable to transplantation. Finally, transplantation of xeno-free vmDA progenitors from LMX1A- and PITX3-eGFP reporter lines into Parkinsonian rodents demonstrate improved engraftment outcomes and restoration of motor deficits.
Adult SVZ Stem Cells Lie in a Vascular Niche: A Quantitative Analysis of Niche Cell-Cell Interactions
Shen Q., Wang Y., Kokovay E., Lin G., Chuang S-M. Goderie S.K., Roysam B., Temple S. Cell Stem Cell, 2008
Here, they examine the relationship of adult SVZ NSC lineage cells to blood vessels using confocal imaging of SVZ whole mounts in which the normal 3D relationships of cells are preserved. Neural stem cells (NSCs) in the adult subventricular zone (SVZ) lie close to blood vessels in a vascular-derived laminin-rich niche. Cells expressing stem cell markers, including GFAP, and proliferation markers are closely apposed to the laminin-containing extracellular matrix (ECM) surrounding vascular endothelial cells. Adult SVZ progenitor cells express the laminin receptor a6B1 integrin, and blocking this inhibits their adhesion to endothelial cells, altering their position and proliferation in vivo.
