Maulana et al. (2024) Solid tumor-on-chip model for efficacy and safety assessment of CAR-T cell therapy. Cell Stem Cell 31, 1-14 (Article)

Human breast cancer cells were embedded in 3-D Life Hydrogel on a novel microfluidic system to assess its applicability in investigating CAR-T cell mediated therapeutic treatments. The publication shows how CAR-T cells can migrate into the hydrogel towards the embedded tumor cell aggregates and how subsequent cytokine release can be measured. It is also shown how the chip-based hydrogel cell culture can be used to assess treatments for antagonizing the Cytokine Release Syndrome. Cells used: MDA-MB-231 (human breast cancer cell line); primary human fibroblasts, human breast cancer organoids (patient derived), human T cells, CAR-T cells. Methods used: Injection of 3-D Life Hydrogel into microchannels; fluorescence microscopy.

Chen et al. (2023) Enteric neurospheres retain the capacity to assemble neural networks with motile and metamorphic gliocytes and ganglia. Stem Cell Research & Therapy 14:290 (Article)

Enteric neurospheres assemble enteric nervous system networks in 3-D Life ToGro Hydrogel. Cells used: murine enteric neural stem cells (neurospheres). Methods used: microscopy of neural networks.

Militaru et al. (2023) New panel of biomarkers to discriminate between amelanotic and melanotic metastatic melanoma. Front. Oncol. 12:1061832 (Article)

3-D Life Hydrogel was used to perform a migration assay on different melanoma cell lines to compare cell motility and evaluate their cell migration capacity. A small drop of Dextran-PEG hydrogel was placed in wells of a 96 well plate and cells were seeded around the gel drop. After 24 hours the gel was dissolved by 3-D Life Dextranse leaving cells intact and attached to the bottom of the well. Cell migration into the empty space left by the dissolution of the gel was subsequently recorded and quantified by image analysis.

Kromidas et al. (2023) Immunocompetent PDMS-Free Organ-on-Chip Model of Cervical Cancer Integrating Patient-Specific Cervical Fibroblasts and Neutrophils. Adv. Healthcare Mater.:2302714 (Link)

The 3-D Life Hydrogel system was used in a microfluidic platform specifically developed for the 3-D co-cultivation of human cervical cancerous tissue (spheroids) with donor-derived cervical fibroblasts. Donor-derived neutrophils were integrated in the microfluidic system and were recruited into the hydrogel-based co-culture. The 3-D Life hydrogel demonstrated good stability without shrinkage and provided a robust 3D matrix for the cells. The treatment of the culture with cisplatin demonstrates the applicability of the platform for drug testing and the development of new (immuno) therapeutic options. Application: Hydrogel in a 3D microfluidic platform for drug discovery. Cells used: SiHa (human squamous cervical cancer cell line), primary fibroblasts (from human cervix uteri), human PMNs (polymorphonuclear leukocytes/neutrophils from whole blood), human PBMCs (peripheral blood mononucleated cells from whole blood). Methods used: Life/Dead staining (Calcein AM/PI), immunofluorescent staining, LDH assay (cytotoxicity), TUNEL Assay, Live cell tracking in the hydrogel.

Hafa et al. (2023) Laser patterning bioprinting using a light sheet-based system equipped with light sheet imaging produces long-term viable skin constructs. Adv. Mater.: e2306258 (Article)

Newly developed light-induced hydrogels of Cellendes (product in development) were used in a novel integrated fluorescent light sheet bioprinting and imaging system that combines high printing speed and high resolution with light sheet-based imaging. A resolution of 9 µm was achieved in printed structures which was, up to date, only reached by two photon photopolymerization. No hydrogel contraction or expansion through the bioprinting procedure was observed. Using this bioprinting system full-thickness skin constructs were generated and remained viable for 41 days. Cells used: HaCaT (human keratinocytes), Hs27 (human fibroblasts), HUVEC. Methods: diffusion through hydrogel by Fluorescence Recovery after Photobleaching (FRAP) analysis; light sheet based bioprinting; immunofluorescence staining.

Krüger et al. (2022) Sensitizer-enhanced two-photon patterning of biomolecules in photoinstructive hydrogels. Communications materials 3 (9) (Article)

Using two-photon microscopy the authors developed a method to tether proteins or peptides in the hydrogel network with micrometer precision. This allows to follow the interaction and effects of immobilised proteins or peptides with cells in subcellular dimensions.

Del Mistro et al. (2022) Focal adhesion kinase plays a dual role in TRAIL resistance and metastatic outgrowth of malignant melanoma. Cell Death and Disease 13:54 (Article)

Cipriano et al. (2022) Human immunocompetent choroid-on-chip: a novel tool for studying ocular effects of biological drugs. Communication Biology 5: 52 (Article)

3-D Life Hydrogel was used in a microphysiological system (MPS) to build the stromal compartment of a human choroidal in vitro model (Choroid-on-Chip). The model allowed for a controlled immune cell recruitment into the melanocyte-bearing stromal compartment through a vascular monolayer. This choroid-on-chip model was shown to effectively work in efficacy testing of immunosuppressive compounds as well as safety profiling of immunoactivating antibodies. Cells used: primary human melanocytes. Methods used: microphysiological system, perfusion, live/dead staining, KI-67 staining, nuclear (DAPI) staining, antibody staining, confocal microscopy and analysis.

Albrecht et al. (2022) Mechanistically Coupled PK (MCPK) Model to Describe Enzyme Induction and Occupancy Dependent DDI of Dabrafenib Metabolism. Pharmaceutics 14: 310 (Article)

Hensler et al. (2021) A novel standardized inflammatory cell-modulated 3D tumor tissue model for analysis of tumor-stroma interaction and drug discovery. American Journal of Bioscience and Bioengineering 9(4): 110-122 (Link)

Using the 3-D Life Hydrogel system a reproducible tumor-stroma model including macrophages, neutrophils and fibroblasts within a malignant tumor microenvironment was developed. In contrast to collagen-based matrices, where the matrix strongly contracts during a long culture period, the 3-D Life Hydrogel maintained its size. Application: 3D tumor-stroma model for drug discovery. Cells used: primary human fibroblasts; U937 lymphoblast cell line (differentiated to macrophages), HL-60 promyelocytic cell line (differentiated to neutrophils), HaCaT-ras A-5RT3 (tumor keratinocytes), HaCaT-ras A-5IL-6 (tumor keratinocytes), MCF-7 (mamma carcinoma cells), H838 (lung carcinoma cells). Methods used: hydrogel culture in 24 well transwell insert plates. Fibroblasts and immune cells were cultured in the gel, tumor epithelial cells on top of the gel. Cultures with keratinocytes were cultivated at the air-liquid-interface.

Nair et al. (2021) Parallelizable Microfluidic Platform to Model and Assess In Vitro Cellular Barriers: Technology and Application to Study the Interaction of 3D Tumor Spheroids with Cellular Barriers. Biosensors 11(9), 314 (Article)

3-D Life Hydrogel was used to create an extracellular matrix-lumen interface within a microfluidic chip. The successful co-culture of tumor spheroids in the gel and the epithelial cell layer on the gel surface shows the suitability of the setup as a cellular barrier model. Application: cell-based assays in microfluidic platforms; in vitro model of cellular barriers; transepithelial cell migration. Cells Used: HT29, MDCK. Methods used: hydrogel in a microfluidic platform; transepithelial electrical resistance (TEER) measurements.

Wang et al. (2021) Spatial micro-variation of 3D hydrogel stiffness regulates the biomechanical properties of hMSCs. Biofabrication 13: 035051 (Article)

The 3-D Life Hydrogel system was used to generate hydrogels with controlled variations in local stiffness at microscale dimensions to examine mechanotransductional effects on human mesenchymal stem cells. Application: cellular mechanotransduction. Cells Used: human mesenchymal stem cells (hMSCs). Methods used: atomic force microscopy; rheology; Live/dead staining; immunostaining; paraffin embedment and sectioning; hematoxylin-eosin staining; RT-qPCR.

Lang et al. (2021) Architecture-Promoted Biomechanical Performance-Tuning of Tissue-Engineered Constructs for Biological Intervertebral Disc Replacement. Materials 14, 2692. (Article)

Chondrons prepared from articular cartilage were cultured in 3-D Life Dextran-PEG Hydrogels modified with 3-D Life RGD Peptide in a specific architectural design. The resulting 3D tissue constructs were mechanically characterized. Application: Tissue engineering. Cells Used: chondrons derived from articular cartilage. Methods used: Live-dead staining; compression loading.

Bavli et al. (2021) A highly sensitive analysis platform for the characterization of 3D-cultured single-cell-derived clones. Developmental Cell 56, 1–14. (Link)

Single cells are encapsulated in 3-D Life Hydrogel spheres via an oil-phase ‘‘pinching’’ process in a microfluidic device and subsequently cultured in multiwell plates to generate clones of cells. To culture mouse embryonic stem cells (mESCs) in these spheres the hydrogel components were mixed with several additives including gelatin. mESCs maintained pluripotency over at least 8 days of culture indicating the suitability of this 3D culturing system to promote stemness of ESCs even without pluripotent media. Application: single cell analysis. Cells Used: PC9 (lung adenocarcinoma, human), ZHBTc4 ESCs (mouse embryonic stem cells). Methods used: microsphere production, microsphere cultivation, scRNA-seq, fluorescent labeling of hydrogel via biotin-streptavidin attachment.

Bavli et al. (2021) CloneSeq - Single-cell clonal 3D culture and analysis protocol. STAR Protocols 2, 100794, December 17, 2021 (Article)

Detailed protocol on the generation of microspheres for single cell encapsulation.

Yin et al. (2021) Cell migration regulated by spatially controlled stiffness inside composition-tunable three-dimensional dextran hydrogels. Adv. Mater. Interfaces 8, 2100494 (Link)

The authors studied the regulation of cell migration by tuning the spatial stiffness of hydrogels. Migration velocity and cell morphology were analyzed to charaterize the dependence of cell migration on the heterogeneous stiffness in subcellular scale. Application: mechanotransduction. Cells Used: C2C12 murine myoblasts. Methods used: live/dead cell staining; cell tracking; rheological and atomic force microscopic measurements of hydrogels; scanning electron microscopy of hydrogels; fluorescent staining (phalloidin) and fluorescent antibody staining; fluorescence imaging of cells in gels.

Jung et al. (2021) Non-invasive analysis of pancreas organoids in synthetic hydrogels defines material-cell interactions and luminal composition. Biomaterials Science 9(16):5415-5426 (Article)

Kuehlbach et al. (2021) Recapitulating the Angiogenic Switch in a Hydrogel-Based 3D In Vitro Tumor-Stroma Model. Bioengineering 8: 186 (Article)

Zhao et al. (2021) Intermittent pressure imitating rolling manipulation ameliorates injury in skeletal muscle cells through oxidative stress and lipid metabolism signalling pathways. Gene 778: 145460 (Link)

Palau et al. (2021) Both Specific Endothelial and Proximal Tubular Adam17 Deletion Protect against Diabetic Nephropathy. Int. J. Mol. Sci. 22: 5520 (Article)

Georgakopoulos et al. (2020) Long-term expansion, genomic stability and in vivo safety of adult human pancreas organoids. BMC Developmental Biology 20:4 (Article)

Cellendes hydrogel is used to culture human pancreas organoids in a chemically definded 3D matrix.

Trennheuser et al. (2020) 3D Culture and Characterization of Blood-Brain Barrier Endothelial Cells in a New Microfluidic Platform Research Square (Article)

Brancato et al. (2020) Could 3D models of cancer enhance drug screening? Biomaterials 232: 119744 (Link)

Rothbauer et al. (2020) Monitoring tissue-level remodelling during inflammatory arthritis using a three-dimensional synovium-on-a-chip with non-invasive light scattering biosensing. Lab Chip 20, 1461 (Link)

Friedrich et al. (2019) Stretch in Focus: 2D Inplane Cell Stretch Systems for Studies of Cardiac Mechano-Signaling. Front Bioeng Biotechnol. 27;7:55 (Article)

Murine ventricular cardiomyocytes were embedded in disks of PVA-PEG/RGD hydrogels and placed on a stretching device (IsoStretcher) to examine the conversion of mechanical stimuli into Ca2+ signaling. The cardiomyocytes in the gel were loaded with the Ca2+ indicator Fluo-4 AM and calcium transients were recorded upon radial stretching of the cells.

Kraus et al. (2019) Evaluation of a 3D Human Artificial Lymph Node as Test Model for the Assessment of Immunogenicity of Protein Aggregates. J Pharm Sci.108(7):2358-2366 (Link)

3-D Life Hydrogel is used in an artificial lymph node model using a perfused bioreactor system. PBMCs and stromal cells are cultured in this model.

Zippel et al. (2019) Migration Assay for Leukemic Cells in a 3D Matrix Toward a Chemoattractant. Methods Mol Biol. 2017:97-107 (Link)

The Dextran-CD Hydrogel FG is used for an in vitro 3D chemotaxis assay for leukemic cells. A detailed protocol describes the setup of the assay in µ-Slides Chemotaxis (ibidi GmbH, Munich,Germany), cell tracking and quantitative analysis of the cell movement towards the chemoattractant.

Shen et al. (2019) Identification and integrative analysis of microRNAs and mRNAs involved in proliferation and invasion of pressure‑treated human liver cancer cell lines. Mol Med Rep. 20(1):375-387 (Article)

Liver cancer cells were embedded in a 3-D Life Hydrogel to analyze the effect of pressure on their proliferative, migratory and invasive ability in a 3D environment.

Xue et al. (2019) Matrix stiffness regulates arteriovenous differentiation of endothelial progenitor cells during vasculogenesis in nude mice. Cell Proliferation 52:e12557 (Article)

The work shows how different stiffnesses of 3-D Life Hydrogels can direct the differentiation of endothelial progenitor cells (EPCs) into a venous or arterial phenotype. The gels injected into nude mice support the in vivo formation of functional blood vessels. Application: in vivo vasculogenesis. Cells used: murine endothelial progenitor cells. Methods used: hydrogel injection into mice; paraffin embedment of gels; antibody staining of tissue slices; dissolution of gels with dextranase and subsequent analysis of cells with qPCR and Western blotting.

Wang et al. (2019) Characterization and Analysis of Collective Cellular Behaviors in 3D Dextran Hydrogels with Homogenous and Clustered RGD Compositions. Biomaterials 35(10): 3273–3280. (Article)

The authors took an innovative approach and used the flexibility of the 3-D Life Hydrogel system to design a clustered RGD Peptide microenvironment to study patterns of cell adhesion ligands on cell behavior. Cells Used: NIH–3T3 fibroblasts, C2C12 cells (myoblasts, murine). Methods used: Live/Dead viability assay, bright-field and confocal microscopy, phalloidin staining, DAPI staining.

Rothdiener M. et al. (2018) Human osteoarthritic chondrons outnumber patient‐ and joint‐matched chondrocytes in hydrogel culture—Future application in autologous cell‐based OA cartilage repair? Journal of Tissue Engineering and Regenerative Medicine 12:e1206-e1220 (Link)

The paper shows a six week cultivation of chondrons in 3-D Life Dextran-PEG Hydrogel.

Hellwig, C. et al. (2018) Culture of human neurospheres in 3D scaffolds for developmental neurotoxicity testing. Toxicology in vitro 52:106-115 (Link)

A novel peptide modification of 3-D Life Hydrogel was used to establish a neurosphere outgrowth assay for developmental neurotoxicity compound testing. The work shows how cell migration, differentiation to neurons and formation of neuronal networks is supported by the hydrogel.

Grobe, H. et al. (2018) A Rac1-FMNL2 signaling module affects cell-cell contact formation independent of Cdc42 and membrane protrusions. PLoS One 13:e0194716 (Article)

3-D Life hydrogel supplemented with RGD Peptide was used for long term culture (14 days) of human breast epithelial cells (MCF10A). Successful lumen formation by spheroids was observed with wildtype cells and investigated with mutated cells in this type of cultures.

Huang et al. (2018) Three-dimensional hydrogel is suitable for targeted investigation of amoeboid migration of glioma cells. Mol Med Rep.17:250-256 (Article)

3-D Life Dextran-CD hydrogel modified with RGD Peptide was used to quantitatively evaluate amoeboid versus mesenchymal cell migration of glioma cells. The work shows how 3-D Life Hydrogel, in contrast to 2D culture, supports drug evaluation aiming at the efficient inhibition of both, amoeboid and mesenchymal cell migration in 3D culture. Applications: drug evaluation, cell migration analysis. Methods used: immunofluorescent staining, fluorescence microscopy, chemotaxis in ibidi µ-slides (

He et al. (2018) Cartilage intermediate layer protein is regulated by mechanical stress and affects extracellular matrix synthesis. Molecular Medicine Reports 17: 6130-6137 (Article)

Cartilage intermediate layer protein (CILP) and aggrecan and collagen II synthesis were measured in human nucleus pulposus (NP) cells in response to mechanical stimuli, including cyclic compressive stress and cyclic tensile strain. Application: Effects of mechanical stress on protein expression. Cells used: nucleus pulposus cells. Methods used: NP cells were embedded in cylinders of 3-D Life Dextran-PEG FG hydrogels (Cat. No. G90-1). The hydrogel cylinders were introduced into Bioflex 6-well compression plates and mechanical stress was applied using a FX-5000C™ Flexercell system (Flexcell International). Cells were extracted from hydrogels using 3-D Life Dextranase (Cat. No. D10-1) and subjected to RT-PCR and Western blotting analysis.

Miyakawa et al. (2018) Development of a cell-based assay to identify hepatitis B virus entry inhibitors targeting the sodium taurocholate cotransporting polypeptide. Oncotarget 9(34), 23681-23694 (Article)

Spheroids of HepG2 cells were grown in 3-D Life Dextran-CD Hydrogels. Application: spheroid culture. Cells Used: HepG2. Methods used: Immunofluorescence, confocal microscopy.

Friedrich, O. et al. (2017) Adding dimension to cellular mechanotransduction: Advances in biomedical engineering of multiaxial cell-stretch systems and their application to cardiovascular biomechanics and mechano-signaling. Progress in Biophysics and Molecular Biology 130:170-191 (Link)

The effect of multiaxial cell stretch on adult ventricular cardiomyocytes (CM) embedded in 3-D Life Cellendes hydrogel of different stiffnesses (Young modulus of 1 kPa, 4-9 kPa and > than 10 kPa) was examined on a cell stretch system (IsoStretcher). Single CMs were embedded in SG-PVA-PEG hydrogels modified with RGD Peptide to allow for adhesion of cells to the surrounding hydrogel. Stretch-induced Ca2+ influx was measured with the Ca2+ indicator Fluo-4 AM which was mixed into the hydrogel before gelation.

Ayenehdeh et al. (2017) Immunomodulatory and protective effects of adipose tissue-derived mesenchymal stem cells in an allograft islet composite transplantation for experimental autoimmune type 1 diabetes. Immunol Lett.188:21-31 (Link)

3-D Life Hydrogel was used to co-culture pancreatic islets with adipose tissue derived mesenchymal stem cells (AT-MSCs) to determine the effect of AT-MSCs on islet insulin secretion. Hydrogel cultures were examined in vitro to determine insulin secretion as well as transplanted in diabetic mice to determine the effect on blood glucose levels. The implanted hydrogel prevented passage of immune cells to the allograft, as shown after recovery of the implant and subsequent processing for histopathological examinations. Islets and cells were recovered from explanted hydrogels by dextranase treatment and subjected to RT-PCR analyses.

Angres, B. and Wurst, H. (2017) 3-D Life biomimetic hydrogels: A modular system for cell environment design. Przyborski, S. (ed.) Technology Platforms for 3D Cell Culture, A User`s Guide. pp. 197-221. (Link)

This book chapter describes in detail the 3-D Life Hydrogel platform technology as well as selected applications of in vitro cultures.

Nugraha et al. (2017) Monitoring and manipulating cellular crosstalk during kidney fibrosis inside a 3D in vitro co-culture. Sci Rep. 7:14490 (Article)

A disease model of kidney fibrosis was developed using 3-D Life Dextran hydrogel by co-culturing human kidney epithelial cells and human fibroblasts in two layers of hydrogels of different biomimetic modifications. The work shows a unique and successful model system for screening of new molecules capable to interfere and modulate the dialogue between epithelial and mesenchymal cells. Application: target identification and drug evaluation. Methods used: immunofluorescent staining and fluorescence imaging, drug administration, transmission electron microscopy, proliferation assay, RNA extraction for gene expression array.

Noguchi et al. (2017) Molecular analysis of keratocystic odontogenic tumor cell lines derived from sporadic and basal cell nevus syndrome patients. Int. J. Oncol. 51:1731-1738 (Article)

The authors established two keratocystic odontogenic tumor (KCOT) cell lines which they cultured in PVA-PEG-based 3-D Life Hydrogels modified with RGD Peptide. Stainings of the actin cytoskeleton with rhodamine phalloidin and nuclei with DAPI showed spheroid formation with different characteristics depending on the patient's disease and origin of the KCOT cell line.

Weyel et al. (2017) A Two-Photon-Photocleavable Linker for Triggering Light-Induced Strand Breaks in Oligonucleotides. ACS Chem. Biol. 12: 2183-2190 (Link)

Sardi, M. et al. (2016) Modeling Human Immunity In Vitro: Improving artificial lymph node physiology by stromal cells. Appl Vitr Toxicol. 2016:2(3);143-150. (link)

3-D Life Hydrogel is used to develop an artificial lymph node model using a perfused bioreactor system. PBMCs, antigen-presenting dendritic cells, and MSC-derived stromal cells are co-cultivated.

Koenig, G., et al. (2016) Cell-laden hydrogel/titanium microhybrids: Site-specific cell delivery to metallic implants for improved integration. Acta Biomater. 2016 Mar;33:301-10. doi: 10.1016/j.actbio.2016.01.023. Epub 2016 Jan 21. (link)

Co-culture of HUVECs and fibroblasts in 3-D Life Hydrogels to assess titanium-hydrogel-cell compatibility for future implantation strategies.

Grikscheit, K. et al. (2015) Junctional actin assembly is mediated by Formin-like 2 downstream of Rac 1. J. Cell Biol. 209:367-76. (PubMed)

Molecular mechanisms of de novo epithelial lumen formation is studied in long term cultures of MCF10A mammary epithelial cells in 3-D Life Hydrogels.

Charwat, V. et al. (2015) Potential and limitations of microscopy and Raman spectroscopy for live-cell analysis of 3D cell cultures. J Biotechnol. 205:70-81 (link)

Cancer cells and fibroblasts are analyzed alone and in co-culture in 3-D Life Hydrogels using Raman spectroscopy.

Sun, J. et al. (2014) Geometric control of capillary architecture via cell-matrix mechanical interactions. Biomaterials. 35:3273-80. (Article)

3-D Life Dextran-PEG Hydrogel was mixed with Matrigel to adjust the stiffness of Matrigel while maintaining the ligand density for cell adhesion.

Rimann, M. et al. (2014) Automation of 3D Cell Culture Using Chemically Defined Hydrogels. J. Lab. Autom. 19:191-197 (link)

Automated drug screening of tumor spheroids in 3-D Life Hydrogels. Demonstrates the different drug sensitvities of tumor cells in 2-D versus 3-D cell culture.

Ueda, E., et al. (2012) DropletMicroArray:Facile Formation of Arrays of Microdroplets and Hydrogels Micropads for Cell Screening Applications. Lab Chip 12:5218-5224 (PubMed)

Preparation of hydrogel microarrays with 3-D Life Hydrogel for research and high-throughput screening.

Rimann, M., Graf-Hausner, U. (2012) Synthetic 3D Multicellular Systems for Drug Development. Curr. Opin. Biotechnol. 23:803-809 (PubMed)

Review on synthetic 3D culture systems, including 3-D Life Hydrogel.

Neugebauer, U., et al. (2012) From Infection to Detection: Imaging S aureus-host Interactions. Biomed. Tech. (Berl) (link)

3-D Life Hydrogel is used to immobilize bacteria for Raman spectroscopy.

Benz, K., et al. (2010) Polyethylene Glycol-Crosslinked Serum Albumin/Hyaluronan Hydrogel for the Cultivation of Chondrogenic Cell Types. A Adv. Eng. Mater. 12:B539-B551 (link)

3-D Life Hydrogel technology is used with maleimide-modified serum albumin for the cultivation of chondrogenic cells.

Scholz, B., et al. (2010) Suppression of Adverse Angiogenesis in an Albumin-based Hydrogel for Articular Cartilage and Intervertebral Disc Regeneration. Eur. Cell. Mater. 20:24-37 (download)

3-D Life Hydrogel technology used with maleimide-modified serum albumin.

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