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.
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.
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.
Generation of 3D retinal tissue from human pluripotent stem cells using a directed small molecule-based serum-free microwell platform
Hassan Rashidi, Yeh Chwan Leong, Kerrie Venner, Hema Pramod, Qi-Zhen Fei, Owen J. R. Jones, Dale Moulding & Jane C. Sowden Scientific Reports, 2022
Biolaminin 521 (LN521) was used as the matrix in a 2D/3D system for retinal differentiation from human pluripotent stem cells in an agarose micromould platform. To achieve a serum-free and animal-free protocol for producing retinal tissue with photoreceptor cells, the scientists successfully replaced Matrigel (which is derived from mouse tumor cells) with laminin-521, and fetal bovine serum (FBS) with human platelet lysate (HPL). The culture system first allows cells to self-organize forming neuroepithelial structures that resemble embryonic optic vesicles, and then mimics retinogenesis and produces retinal tissue with photoreceptor cells. The generated photoreceptors exhibited key photoreceptor cell features including the formation of OS-like structures containing proteins involved in phototransduction.
Toward Xeno-Free Differentiation of Human Induced Pluripotent Stem Cell-Derived Small Intestinal Epithelial Cells
Jaakko Saari, Fatima Siddique, Sanna Korpela, Elina Mäntylä, Teemu O. Ihalainen, Katri Kaukinen, Katriina Aalto-Setälä, Katri Lindfors, and Kati Juuti-Uusitalo. International Journal of Molecular Sciences, 2022
Small intestinal epithelial cells were differentiated from induced pluripotent stem cells (iPSC-SIEC), via definitive endoderm, on Biolaminin 511. The aim of the study was to develop a protocol with animal-free components to reduce variation in experimental outcomes. The differentiated cells cultured on laminin exhibited more enterocyte specific cellular functionality than cells on Geltrex or collagen. The protocol is serum-free and can be easily modified as completely xeno-free by culturing iPSCs on Biolaminin 521 matrix and changing the B27 and N2 supplement to available xeno-free versions.
Pluripotent stem cell-derived cardiovascular progenitors differentiated on laminin 221 regenerate and improve function of infarcted swine hearts
Yap L, Chong LY, Tan C, Adusumalli S, Seow M, Guo J, Cai Z, Loo SJ, Lim E, Lath N, Ye L, Petretto E, Tryggvason K. BiorXiv, 2021
The authors report a highly reproducible, chemically defined and fully humanized differentiation method of hESCs for the generation of potent cardiovascular progenitors (CVPs), using laminin-221 (Biolaminin 221) as the culture matrix. Transplantation of the CVPs into the myocardial infarcted pig hearts yielded maturation of the progenitor cells to cardiomyocytes and improved cardiac function using only 200 million CVPs, without fatal ventricular arrhythmia occurring. The results may have a significant impact on regenerative cardiology and the method is intended to be used in stem cell therapy of myocardial infarction in humans.
Improved erythroid differentiation of multiple human pluripotent stem cell lines in microcarrier culture by modulation of Wnt/β-Catenin signaling
Jaichandran Sivalingam, Hong Yu Chen, Bin-Xia Yang, Zhong Ri Lim, Alan Tin Lun Lam, Tsung Liang Woo, Allen Kuan-Liang Chen, Shaul Reuveny, Yuin-Han Loh, Steve Kah-Weng Oh. Haematologica, 2018
This article describes means to scale up the pluripotent expansion stage by culturing hiPSCs on Biolaminin 521 (LN521)-coated microcarriers (MCs). The protocol allows hiPSC-MC aggregates to efficiently differentiate differentiate as embryoid bodies (EBs) in a scalable manner in suspension culture.
A Scalable Suspension Platform for Generating High-Density Cultures of Universal Red Blood Cells from Human Induced Pluripotent Stem Cells
Jaichandran Sivalingam, Yu SuE, Zhong Ri Lim, Shaul Reuveny, Benoit Malleret, Steve K.W. Oh. Stem Cells Reports, 2020
This article describes a scalable suspension agitation culture platform for the differentiation of human induced pluripotent stem cell-microcarrier aggregates into functional red blood cells. The report describes the cell quantity and quality in culture sizes ranging from 6-well plates to 500 ml spinner flasks, reaching up to 17 million high-quality cells per ml. The process could find applications in future large-scale red blood cell production in controlled bioreactors.
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.