Create Endothelial Tube Networks in LunaX™ ECM
Summary This protocol describes a 3D endothelial tube formation assay using HUVECs co-cultured with pericytes or mesenchymal stromal cells in LunaX ECM. The model supports mechanistic angiogenesis research, preclinical screening and evaluation of pro- or anti-angiogenic treatments. What you’ll do Prepare LunaX MultiMatrix Low Stiffness ECM and fresh photoinitiator. Combine HUVECs with pericytes or MSCs […]
Paraffin Embedding and Sectioning of LunaX™ Hydrogel Constructs
Summary This protocol adapts standard tissue histology methods for paraffin embedding and sectioning of LunaX hydrogel constructs. It covers fixation, graded dehydration, clearing, paraffin infiltration, embedding and microtome sectioning for downstream histology, special stains and immunohistochemistry. What you’ll do Fix LunaX constructs in 4% paraformaldehyde. Dehydrate samples through a graded ethanol series. Clear constructs in […]
Stain and Image Cryosectioned LunaX™ Hydrogel Constructs
Summary This protocol covers OCT embedding, cryosectioning, staining and imaging of fixed LunaX hydrogel constructs. It supports histology, histochemistry, immunohistochemistry and immunofluorescence for high-resolution analysis of cellular structures, proteins and extracellular matrix components. What you’ll do Fix LunaX hydrogel constructs and embed them centrally in OCT compound. Prepare approximately 10 µm cryosections on positively charged […]
Whole-Mount Immunofluorescence Staining of LunaX™ Constructs
Summary This protocol describes whole-mount immunofluorescence staining of intact 3D cultures encapsulated in LunaX hydrogels. It supports visualisation of nuclear, cytoskeletal, cell-surface and intracellular targets without sectioning, using widefield or confocal fluorescence microscopy. What you’ll do Fix intact LunaX constructs with 4% paraformaldehyde. Permeabilise and block samples to reduce non-specific antibody binding. Incubate constructs sequentially […]
Differentiate C2C12 Myoblasts into Myotubes in LunaX™ ECM
Summary This protocol provides starting conditions for differentiating C2C12 myoblasts into contractile myotubes within LunaX ECM. The tuneable 3D matrix supports skeletal muscle modelling across physiologically relevant stiffness conditions while helping minimise construct shrinkage during culture. What you’ll do Prepare LunaX MultiMatrix Low Stiffness ECM and fresh photoinitiator. Encapsulate C2C12 cells at a recommended density […]
Run an ECM-Embedded 3D Spheroid Invasion Assay
Summary This protocol measures invasive outgrowth from preformed cancer spheroids embedded within LunaX ECM. The 96-well model surrounds each spheroid with a tuneable 3D matrix for studying stiffness-dependent invasion, treatment response and differences between cancer cell types. What you’ll do Form compact cancer spheroids in an ultra-low-attachment plate. Embed individual spheroids in 30 µL of […]
Run a Surface-Seeded 3D Spheroid Invasion Assay
Summary This protocol quantifies invasive outgrowth from preformed cancer spheroids seeded onto LunaX ECM hydrogels. The 96-well assay supports comparisons of matrix stiffness, treatment effects and intrinsic invasive potential using daily brightfield imaging and image-based quantification. What you’ll do Generate compact spheroids in a 96-well ultra-low-attachment plate. Prepare cell-free LunaX ECM surfaces across a defined […]
Generate Patient-Derived Organoids in LunaX™ ECM
Summary This protocol describes the encapsulation of primary tumour-derived cells in LunaX CoreMatrix Low Stiffness ECM to establish patient-derived organoids. It provides starting conditions for reproducible 30 µL dome cultures in a defined, tuneable 0.5–1.5 kPa matrix. What you’ll do Prepare LunaX ECM precursor and freshly reconstituted photoinitiator. Harvest expanded primary tumour cells and remove […]
Isolate and Expand Primary Tumour Cells from Cancer Tissue
Summary This protocol describes how to isolate viable primary tumour cells from fresh biopsies or resected cancer tissue and expand them to approximately 80% confluence. The resulting cultures provide a physiologically relevant starting point for patient-derived organoids and other LunaX™ 3D models. What you’ll do Prepare and mince fresh cancer tissue under aseptic conditions. Digest […]