3D cell culture using Biolaminin and Biosilk matrices
3D culture systems for disease modeling, drug screening and regenerative medicine
Cell microenvironment consists of extracellular matrix, oxygen and nutrient gradients, and both cell-to-matrix and cell-to-cell interactions. In order to enhance biological research, drug discovery process and regenerative medicine applications, cells are now often cultured in a three dimensional environment (3D) instead of two dimensional monolayer cultures.
Recent advances in stem cell culture have made it possible to grow organoids, in vitro 3D tissues. Organoid protocols have been described for a variety of organs, including digestive tract, kidney, and the brain. Low success rate in clinical drug trials have shown the need for improved preclinical 3D culture systems that accurately represent in vivo conditions and tissue-specific characteristics, allowing drug safety and efficacy assessment from an earlier stage of drug development. 3D models applied to a high-throughput format are under development for large-scale screening systems.
Biolaminin substrates are bioactive components that can be added to a variety of 3D culture systems
Scientists have long been developing more reproducible 3D gel and network models in order to replace the previously commonly used xenogeneic matrix extracts, such as the ones derived from Engelbreth-Holm-Swarm mouse sarcoma tissue. Now scientists agree that these extracted matrices are too undefined and famously suffer from high batch-to-batch variability and xenogeneic origin, preventing robust protocols and clinical use. To accommodate these needs, Biolaminin and Biosilk products are animal origin free and documented even for clinical use, in addition to giving prominent advantages for cell culture applications.
Full-length laminins have proven to be crucial components for proper organoid formation in gel models. For example, Gjorevski and co-authors (2016) showed that only the full-length laminin-111, and not laminin-derived peptides, is sufficient for intestinal organoid formation in a defined hydrogel matrix. By using different Biolaminin products, many culture systems can also be fine-tuned to meet the tissue-specific characteristics of an in vivo-like microenvironment.
Dobre and co-authors (2021) developed 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 native, 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. The study appreciated the tissue-specific interactions between full-length laminin isoforms and growth factors – laminins do not only support cell function by interacting directly, but also by binding with large range of growth factors and presenting them effectively to the cells. This relatively recently discovered property makes laminins even more important components for tissue engineering.
Ye and coauthors (2020) reported a defined, nonimmunogenic hydrogel matrix utilizing Biolaminin-111 as a bioactive component for liver organoid culture. The authors show that LEC, which is derived from mouse sarcoma, can be replaced with the human recombinant Biolaminin-111 resulting in similar proliferative potential, which makes the hydrogel system completely synthetic for the expansion of human liver organoids potentially even for clinical applications.
An economical and automated platform for the generation of human liver spheres was developed by Lucendo-Villarin and co-authors (2020). Stem cell derived liver tissues with hepatic progenitors, endothelial cells and hepatic stellate cells, differentiated with the help of Biolaminin 521, permit the study of human liver biology and can be scaled for drug screening. A detailed protocol for this effective model system was also published (Meseguer-Ripolles et al. 2021).
Biosilk is an effective and unique tool for organoid development
Biosilk is a natural biomaterial that has the ability to self-assemble into a microfibrous network, such as foams, and can easily be biofunctionalized with different ECM proteins like laminin proteins, to better recapitulate the more physiologically relevant aspects of developing human tissue. Recombinant human laminin 521, Biolaminin 521 has been shown advantageous in promoting self-renewal of high-quality human pluripotent stem cells (hPSCs). The combination of Biosilk with Biolaminin 521 (Biosilk 521) is an ideal 3D substrate for hPSC integration and expansion in the foam form of the 3D Biosilk scaffold. Lifting the foams from plate bottom promotes the following long-term differentiation as free-floating cerebral organoids. The hPSCs rapidly expand with the same expression of pluripotency markers as the 2D control and after a few days, sufficient cell density of a homogenous, pluripotent starting cell population has been achieved for initiation of long-term, in situ differentiation towards neural lineages (Åstrand, 2020). To support cells that reside in different tissue microenvironments in vivo, other Biolaminin substrates can be easily added to the Biosilk culture system.
The fibrillar Biosilk network allows the formation of channels throughout the 3D culture, which facilitates diffusion of oxygen, medium, and patterning factors (Åstrand et al. 2020). This enables long-term differentiation protocols and makes it possible to generate larger organoids with uniform cellular specialization and organization, without an increased risk of getting necrotic centers. Beside these advantages, Biosilk is also biodegradable and non-immunogenic biomaterial, which facilitates the future use in clinical applications. These advantages make Biosilk a great choice for successful 3D applications.
Succeed with your application
Instructions: Biosilk 3D scaffolds for differentiation and proliferation of hPSCs
Protocol for human pluripotent stem cell expansion, differentiation and organoid formation in 3D
Assembly of FN-silk with laminin-521 to integrate hPSCs into a three-dimensional culture for neural differentiation
Carolina Åstrand, Veronique Chotteau, Anna Falk, My Hedhammar Biomater Sci, 2020
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
Guide: A workflow for preparation of 3D Biosilk breast cancer model
Biosilk pipetting optimization with Sartorius Picus Nxt pipette
Application note: Biosilk 3D biomaterial for organoid culture
Features and supporting data for Biosilk in 3D cell culture
Biolaminin and Biosilk Key Advantages
Biosilk has the ability to self-assemble into a microfibrous network, such as foams. Due to its favorable functional and mechanical properties, Biosilk is able to support extensive cellular remodeling, self-organization, and morphogenesis.
In the Biosilk fibrillar network, nutrients can flow into the inner part of the organoid, giving more effective and uniform cellular specialization and organization. Cells do not need to be encapsulated.
Biosilk is a completely defined recombinant spider silk protein matrix which is non-immunogenic and biodegradable, further facilitating the use in clinical applications.
Biolaminin isoforms can be added into different organoid culture systems to support correct cell phenotype. Biosilk 3D network can be easily supplemented with Biolaminin isoforms and other ECM proteins.
All our matrices are chemically defined and animal origin-free, which makes them ideal substrates for each level of the scientific process – from basic research to clinical applications.
3D culture substrateBiosilk is a natural biomaterial made from recombinant spider silk protein, a useful tool for a wide range of 3D culture applications, such as organoid culture and other tissue engineering applications. Biosilk can be mixed with any Biolaminin matrix.VIEW product
3D culture substrateBiosilk 521 is a natural biomaterial made of spider silk and laminin 521 – an ideal 3D culture substrate for integration, expansion, and long-term differentiation of human pluripotent stem cells and different progenitor cells.VIEW product
Talk to our team to get a custom proposal
We are here to help your journey.