Here is a selection of publications where different laminin isoforms were used to create more authentic cell culture systems.
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.
Protocol 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.
CRISPRi-mediated transcriptional silencing in iPSCs for the study of human brain development
Pia Annette Johansson, Anita Adami, Johan Jakobsson. STAR Protocols, 2022
This protocol describes using CRISPRi-mediated transcriptional silencing in human induced pluripotent stem cells. In addition, it contains an efficient protocol for neural progenitor differentiation. The method is directly applicable to loss-of-function studies in brain development research. Biolaminin 521 (laminin-521) is applied as the cell culture matrix for PSCs and Biolaminin 111 (laminin-111) for hiPSC differentiation into forebrain neural progenitor cells. The protocol is expected to achieve high transduction and silencing efficiency. After two weeks of differentiation, the authors detected no reduction in the percentage of positive cells, and the cerebral organoids had stable transcriptional silencing even after 4 months.
Generation of a CHIP isogenic human iPSC-derived cortical neuron model for functional proteomics
Catarina Dias, Erisa Nita. Jakub Faktor, Lenka Hernychova, Tilo Kunath, Kathryn L. Ball. STAR Protocols, 2022
This protocol describes the production of gene-edited cells using CRISPR-Cas9 and a patient-derived induced pluripotent stem cell (iPSC) line. Biolaminin 521 was used as the matrix to improve single cell survival and pluripotency. The resulting panel of iPSC lines was differentiated into cortical neurons with the Biolaminin 111 culture matrix. The overall aim was to identify protein and pathway targets for the neuroprotective E3-ubiquitin ligase CHIP, which is important in healthy brain aging. The protocol can be adapted to other proteins and pluripotent stem cell lines.
Methods for Automated Single Cell Isolation and Sub-Cloning of Human Pluripotent Stem Cells
Valeria Fernandez Vallone, Narasimha Swamy Telugu, Iris Fischer,Duncan Miller, Sandra Schommer, Sebastian Diecke, Harald Stachelscheid. Current Protocols in Stem Cell Biology, 2020
This publication describes automated workflows to facilitate high-throughput hPSC clonal selection and expansion, which is required for example after genome editing. Three different automated single cell dispensing devices were applied with Biolaminin 521 coating of plates and multiwells. Laminin-521 increases single cell cloning efficiency compared to other commonly used matrices. Single cell−derived hPSC sub-clones from each system maintained pluripotency and genetic stability.
Astrocytic laminin-211 drives disseminated breast tumor cell dormancy in brain
Jinxiang Dai, Patrick J. Cimino, Kenneth H. Gouin III, Candice A. Grzelak, Alexander Barrett, Andrea R. Lim, Annalyssa Long, Stephanie Weaver, Lindsey T. Saldin, Aiyedun Uzamere, Vera Schulte, Nigel Clegg, Laura Pisarsky, David Lyden, Mina J. Bissell, Simon Knott, Alana L. Welm, Jason H. Bielas, Kirk C. Hansen, Frank Winkler, Eric C. Holland and Cyrus M. Ghajar. Nature Cancer, 2022
This study shows that astrocyte-deposited laminin-211 drives disseminated tumor cell (DTC) quiescence in brain metastases by inducing the dystroglycan receptor. Dormancy has a key role in the metastasis of breast cancer cells to the brain. The authors compared the laminin 211 isoform with laminin 411, 511, and 111, and only 211 significantly and substantially reduced breast cancer cell outgrowth in addback experiments.
Laminin 511 and WNT signalling sustain prolonged expansion of hiPSC-derived hippocampal progenitors
Keagan Dunville, Fabrizio Tonelli, Elena Novelli, Azzurra Codino, Verediana Massa, Anna Maria Frontino, Silvia Galfrè, Francesca Biondi, Stefano Gustincich, Matteo Caleo , Luca Pandolfini, Claudia Alia, and Federico Cremisi. Development, 2022
The authors identify laminin-511 as a crucial laminin isoform for prolonging the neural stem cell (NSC) state and extending hippocampal NSC proliferation for over 200 days in vitro. Biolaminin 511 supported adhesion and cell cycle progression of the dividing hippocampal progenitors. LN511 was crucial in supporting progenitor proliferation, inhibiting differentiation, and sustaining a gene expression profile responsible for maintaining a hippocampal neurogenic niche for extended periods compared with isoforms LN121, LN332, LN441, and with a mouse laminin product. The study involved a novel protocol for differentiating hippocampal NPCs from human induced pluripotent stem cells via a WNT actuator. The differentiation capability of both young and older NPC populations was retained when tested by xenografting into mice.
Structural and functional polarisation of human pancreatic beta cells in islets from organ donors with and without type 2 diabetes
Louise Cottle, Wan Jun Gan, Ian Gilroy, Jaswinder S. Samra, Anthony J. Gill, Thomas Loudovaris, Helen E. Thomas, Wayne J. Hawthorne, Melkam A. Kebede, and Peter Thorn. Diabetologia, 2021
Isolated human pancreatic beta cells were cultured on Biolaminin 511 -coated glass coverslips to study beta cells' structural and functional polarization. The study shows that human beta cells have a consistent orientation with respect to islet capillaries, surrounded by an extracellular matrix including laminin proteins. Live-cell imaging showed the distribution of insulin granule fusion around the cells. The study recognized beta cell polarity in the native human islet environment which plays a role in insulin secretion.
Chemically defined and xeno-free culture condition for human extended pluripotent stem cells
Bei Liu, Shi Chen, Yaxing Xu, Yulin Lyu, Jinlin Wang, Yuanyuan Du, Yongcheng Sun, Heming Liu, Haoying Zhou, Weifeng Lai, Anqi Xue, Ming Yin, Cheng Li, Yun Bai, Jun Xu & Hongkui Deng. Nature Communications, 2021
This study shows the significant benefit of culturing and deriving extended pluripotent stem cells on Biolaminin 521 (LN521) compared to using Matrigel, Geltrex, vitronectin, extracted laminin, collagen, fibronectin, or laminin-511. In addition to enabling xeno-free and chemically defined culture conditions, the study clearly shows that laminin-521 promotes attachment (measured at 1.5 h), survival (24h), and proliferation (72h). Human EPS cells were long-term and genetically stably maintained in vitro, preserving their embryonic and extraembryonic developmental potentials. The study also showed efficient derivation from human fibroblast through reprogramming.
Enhanced xeno-free differentiation of hiPSC-derived astroglia applied in a blood–brain barrier model
Louise Delsing, Therése Kallur, Henrik Zetterberg, Ryan Hicks, Jane Synnergren. Fluids Barriers CNS, 2019
This study shows that astroglia cells differentiated on Biolaminin 521 display an improved phenotype compared to a mouse EHS-extracted laminin L2020 product. Especially, cells differentiated on Biolaminin 521 had a higher secretion of factors important for BBB formation, such as GFAP, S100B, and Angiopoietin-1, than cells differentiated on the laminin extract. In addition, glutamate uptake and the ability to induce the expression of junction proteins in endothelial cells were affected by the culture matrix choice. The study showed differentiation of functional astroglia from three different human induced pluripotent stem cell lines which were used in a blood-brain barrier model.