Functional Hydrogel Guide

Biomaterials are increasingly being used to study tissue development, tissue engineering, disease pathologies, and regenerative processes. For years, the examination of the biological phenomenon has been routinely explored in two-dimensional (2D) environments on unphysiologically rigid materials, such as glass coverslips. This resulted in a misunderstanding of many cellular behaviors, such as cell shape, proliferation, and differentiation. This 2D system also prevents the proper examination of cellular responses to extracellular cues. In 2D cultures, cells sense their surroundings via dynamic leading-edge filopodia1. However, it is now well known that cells also possess actin-rich apical and basal protrusions, such as invadopodia and podosomes, to sense and respond to the environment in 3D2–4. Invadopodia are critical regulators of extracellular matrix (ECM) remodeling during cancer metastasis, illustrating the importance of studying cellular behaviors in 3D. Hydrogels allow experimenters to encapsulate cells within the hydrogels or seed cells post-formation5. Organic microenvironments, e.g. collagen, expose cells to native culture systems possessing a variety of functional ligands6. Providing cells with a tissue-like environment permits the examination of unique cellular characteristics dependent upon the expression of specific cell surface receptors. However, while organic hydrogels produce a cell culture system similar to in vivo conditions, there are drawbacks5. Using native ECM in cell culture is restrictive due to inconsistencies during production, presence of growth factors, and inconsistencies of ligand bioactivity. This has been overcome via the development of synthetic hydrogels, which are modifiable to improve cell attachment and viability7–9. Synthetic hydrogels permit greater reliability and control of experimental conditions. With greater hydrogel reproducibility, scientists can be assured observed cellular behaviors and characteristics are consistent and accurate.

VitroGel® xeno-free tunable hydrogels closely mimic the in vivo ECM and provide researchers with a highly consistent 3D hydrogel cell culture system10,11. This system offers many advantages over native ECM hydrogels and allows scientists to study biological activity both in vitro and in vivo. In unmodified VitroGel 3D hydrogels researchers can investigate 3D cell culture conditions for cells that require little cell-matrix interaction. Moreover, VitroGel 3D provides the necessary scaffold to promote cell spheroid formation12. In addition to increased consistency, VitroGel 3D hydrogels have been modified with functional ligands to create a well-defined system for 3D cell culture and other applications. For example, VitroGel RGD is a 3D hydrogel modified with immobilized RGD peptide13–17 RGD is an ECM derived cell adhesive peptide that promotes cell attachment and cell-matrix interaction. RGD has been extensively used as an adhesive ligand for several cell types18–27. While RGD is a commonly used adhesive peptide for a wide variety of cell types, VitroGel offers alternative adhesive ligands with more specialized bioactivity. VitroGel COL is a 3D tunable hydrogel system that with an integrin-binding collagen motif, GFOGER. GFOGER was discovered to be critical for collagen and integrin α2β1 binding and has subsequently been used to study cell-collagen matrix interactions. GFOGER modified hydrogels have largely been utilized to study integrin-dependent adhesion28,29, osteogenic differentiation30–33 and cancer cell growth and migration34,35. Alternatively, VitroGel IKVAV and VitroGel YIGSR are modified tunable hydrogels with incorporated functional ligands derived from the laminin α‐1 chain and β1 chain, respectively. IKVAV is used regularly to promote cell adhesion36–39 as well tumor growth35,40, neuronal differentiation9,38,41, neurite outgrowth42,43, and angiogenesis44. While extensively studied as a soluble tumor suppressor in vivo 45–47, due to laminin-binding competition, YIGSR has been additionally used as an immobilized, adhesive ligand for cell culture8,48–50. YIGSR has also been shown to promote endothelial cell differentiation51 and migration52. Due to the unique characteristics of each adhesive peptide, several studies have examined the synergistic effects of RGD, IKVAV, and YIGSR34,53–56. Therefore, different combinations of these bioactive peptides have been incorporated into VitroGel 3D hydrogels, VitroGel LDP1 (RGD+IKVAV+YIGSR), VitroGel LDP2 (RGD+YIGSR), and VitroGel LDP3 (RGD+IKVAV). These different combinations allow researchers to study cellular behaviors in a more complex and ECM-like cell culture system. Finally, in addition to adhesion, ECM remodeling plays a critical role in regulating cell morphology, growth, and mobility in vivo 57. Therefore, the development of cleavable 3D hydrogels has been crucial to understanding tissue development and, perhaps most importantly, cancer cell growth and metastasis58. By incorporating matrix metalloproteases (MMP) cleavable substrates into VitroGel® 3D hydrogels, VitroGel MMP offers scientists the ability to examine MMP regulated cellular behaviors, such as cell proliferation59, migration59–61, invasion62,63, and differentiation64,65, that are regulated by ECM breakdown and remodeling. Importantly, all these different functional modifications are offered in the same xeno-free tunable hydrogel 3D culture system, allowing scientists to investigate different combinations of VitroGel products to find the right conditions for their experiments. This key feature permits examination of cellular responses in a more physiologically relevant context, while still giving experimenters full control. Furthermore, scientists can vary concentrations of individual peptides to find the best conditions for the cells being tested. This is important because cells do no grow and migrate in homogeneous environments66. The ECM consists of around 300 molecules and recapitulating the exact microenvironments would not be feasible67. However, with the VitroGel hydrogel system scientists can combine different functional adhesive ligands, such as VitroGel COL with VitroGel RGD, VitroGel IKVAV, VitroGel YIGSR, or VitroGel LDP1/2/3, to form a heterogeneous, and realistic, microenvironment. Also, it is well known that metastatic cells degrade the local ECM to control invasion and dissemination into new tissue57,68. This process is dependent on both MMP degradation and cell-matrix interaction. By combining VitroGel MMP with VitroGel COL, VitroGel RGD, or Laminin modified hydrogels, the molecular underpinnings of these processes may be examined in a controlled 3D system. Due to the diversity of VitroGel products, a wide variety of cell types from different tissues can be tested.

Cell Behaviors in VitroGel System

Tissue/Organ typeCell TypeRelate productBehavior
Beta cellBL5 human beta cellsVitroGel 3DEnhance spheroids and cluster formation and promote cell viability.
Beta TC3 cellsVitroGel RGDCell proliferation and cellular interations
BoneBone marrow stromal cells (rat)VitroGel RGDOsteogenesic differentiation
VitroGel RGDCell proliferation, cell viability, and cellular networking

Expand Full Table To View All Cell Type Behaviors for All VitroGel Products

Expand To View All Behaviors Table
Tissue/Organ typeCell TypeRelate productBehavior
Beta cellBL5 human beta cellsVitroGel Hydrogel Matrix, VitroGel 3DEnhance spheroids and cluster formation and promote cell viability.
Beta TC3 cellsVitroGel Hydrogel Matrix, VitroGel RGDCell proliferation and cellular interations
BoneBone marrow stromal cells (rat)VitroGel Hydrogel Matrix, VitroGel RGDOsteogenesic differentiation
VitroGel Hydrogel Matrix, VitroGel RGDCell proliferation, cell viability, and cellular networking
VitroGel Hydrogel Matrix, VitroGel COLCell attachment and osteoblast differentiation
Bone marrow mesenchymal stem cells (human)VitroGel Hydrogel Matrix, VitroGel COLChondrogenic/osteogenic differentiation
VitroGel Hydrogel Matrix, VitroGel IKVAVAngiogenesis
VitroGel Hydrogel Matrix, VitroGel COLCell spreading, proliferation, and collagen II production
Bone marrow mesenchymal stem cells (goat)VitroGel Hydrogel Matrix, VitroGel RGDOsteogenesic differentiation
Osteoblasts (rat)VitroGel Hydrogel Matrix, VitroGel RGDCell attachment and spreading
Bone marrow stromal cells (bovine)VitroGel Hydrogel Matrix, VitroGel COLCell spreading and osteocalcin expression
BreastMammary gland MCF10AVitroGel Hydrogel Matrix, VitroGel MMPMMP activity in response to TGF-ß1
Mammary epithelium (mouse)VitroGel Hydrogel Matrix, VitroGel COLCell invasion and dissemination
Cancer/tumorHuman colorectal carcinoma HCT 116VitroGel Hydrogel Matrix, VitroGel RGDcell proliferation, cell survival, and intercelluar networking
Huaman colon carcinoma HCT-8VitroGel Hydrogel Matrix, VitroGel RGDCell proliferation and cell matirx interaction
Glioma U87-MGVitroGel Hydrogel Matrix, VitroGel RGDCell spreading and actin stress fiber assembly
VitroGel Hydrogel Matrix, VitroGel RGDCell proliferation and cell matirx interaction
VitroGel Hydrogel Matrix, VitroGel COLCell migration dependent on mechanical force
VitroGel Hydrogel Matrix, VitroGel MMPcell proliferation, spreading, and migration
Primary glioblastom U87VitroGel Hydrogel Matrix, VitroGel RGDcell proliferation and cellular interations
Glioblastoma SF 268VitroGel Hydrogel Matrix, VitroGel RGDCell proliferation and cell matirx interaction
Glioblastoma SF 295VitroGel Hydrogel Matrix, VitroGel RGDCell proliferation and cell matirx interaction
Glioblastoma SNB75VitroGel Hydrogel Matrix, VitroGel RGDCell proliferation and cell matirx interaction
Glioblastoma U-251 MGVitroGel Hydrogel Matrix, VitroGel RGDCell proliferation and cell matirx interaction
Prostate PC3VitroGel Hydrogel Matrix, VitroGel COLCell proliferation and reduced MMP release
VitroGel Hydrogel Matrix, VitroGel IKVAVcell proliferation and invasion
VitroGel Hydrogel Matrix, VitroGel RGDCell proliferation and invasion
VitroGel Hydrogel Matrix, VitroGel COLCell invasion, migration, and spheroid metabolic activity
Prostate LNCaPVitroGel Hydrogel Matrix, VitroGel RGDCell attachment
VitroGel Hydrogel Matrix, VitroGel COLCell proliferation and prostate specific antigen release
Prostate CRPCVitroGel Hydrogel Matrix, VitroGel RGDCell proliferatin and invasion
Prostate DU145VitroGel Hydrogel Matrix, VitroGel RGDCell proliferation and invasion
Melanoma B16F10VitroGel Hydrogel Matrix, VitroGel COLCell migration, invasion, and MMP release
VitroGel Hydrogel Matrix, VitroGel YIGSRCell attachment and spreading
Breast MDA-MB-231VitroGel Hydrogel Matrix, VitroGel MMPCell invasion
VitroGel Hydrogel MatrixCell spreading
VitroGel Hydrogel Matrix, VitroGel RGDCell proliferation, division, migration, and invasion
VitroGel Hydrogel Matrix, VitroGel COLCell spreading and cluster growth
Fibrosarcoma HT1080VitroGel Hydrogel Matrix, VitroGel COLCell infiltration
VitroGel Hydrogel Matrix, VitroGel COLCell attachment
Breast T47D VitroGel Hydrogel Matrix, VitroGel COLForce dependent tubule formation
VitroGel Hydrogel MatrixCell cluster growth
VitroGel Hydrogel Matrix, VitroGel 3DSpheroid formation and proliferation
VitroGel Hydrogel Matrix, VitroGel COLCell cluster growth
Breast 4T1VitroGel Hydrogel Matrix, VitroGel RGDCell proliferation
Breast CTCVitroGel Hydrogel Matrix, VitroGel 3DCell proliferation
Breast E0771VitroGel Hydrogel Matrix, VitroGel RGDCell proliferation, spheroid formation
Breast AU-565VitroGel Hydrogel Matrix, VitroGel RGDCell proliferation, cell matrix interations
Epithelial ovarian OV-MZ-6VitroGel Hydrogel Matrix, VitroGel RGDSpheroid formation and proliferation
Epithelial ovarian SKOV-3VitroGel Hydrogel Matrix, VitroGel RGDSpheroid formation and proliferation
Glioma U373-MGVitroGel Hydrogel Matrix, VitroGel RGDCell adhesion and migration
Rhabdomyosarcoma (human)VitroGel Hydrogel Matrix, VitroGel YIGSRCell attachment and spreading
Melanoma SK-MEL-28VitroGel Hydrogel Matrix, VitroGel IKVAVCell adhesion and proliferation
Melanoma K-1735VitroGel Hydrogel Matrix, VitroGel IKVAVCell invasion
Melanoma A2058VitroGel Hydrogel Matrix, VitroGel IKVAVCollagenolytic activity
Brainstem glioma DIPGVitroGel Hydrogel Matrix, VitroGel 3DCell proliferation and survival
Hela CellsVitroGel Hydrogel Matrix, VitroGel 3DCell proliferation
Colorectal adenocarcinoma DLD-1 cellsVitroGel Hydrogel Matrix, VitroGel RGDCell proliferation and cell matirx interaction
Glioma LRM55VitroGel Hydrogel Matrix, VitroGel IKVAVCell attachment
Melanoma WM239AVitroGel Hydrogel Matrix, VitroGel MMPCell invasion
Melanoma CellsVitroGel Hydrogel Matrix, VitroGel RGDCell proliferation and cell matirx interaction
Insulinoma ins-1 (Rat)VitroGel Hydrogel Matrix, VitroGel RGDCell proliferation and cell matirx interaction
HEK 293VitroGel Hydrogel Matrix, VitroGel RGDCell proliferation and cell matirx interaction
Biphasic synovial sarcoma SYO-1VitroGel Hydrogel Matrix, VitroGel RGDCell proliferation, cell matirx interaction, and cell survival
Fuji CellsVitroGel Hydrogel Matrix, VitroGel RGDCell proliferation and cell matirx interaction
Chordoma CellsVitroGel Hydrogel Matrix, VitroGel 3DCell proliferation
Bone OSA 1777VitroGel Hydrogel Matrix, VitroGel RGDspheroid and cluster formation
Glioma RuGliVitroGel Hydrogel Matrix, VitroGel COLIntegrin dependent cell adhesion
Breast Cancer MCF-7VitroGel Hydrogel Matrix, VitroGel RGDCell proliferation, intercellular connections
VitroGel Hydrogel Matrix, VitroGel COLCell proliferation, morphological changes, MMP expression, and angiogenesis
Liver carcinoma HepG2VitroGel Hydrogel Matrix, VitroGel COLCell viability, growth, and drug resistance
VitroGel Hydrogel Matrix, VitroGel RGDCell proliferation and cell matirx interaction
Human pancreatic cancer PANC-1VitroGel Hydrogel Matrix, VitroGel RGDcell proliferation and cellular interations
Primary breast (human)VitroGel Hydrogel Matrix, VitroGel COLCell invasion, migration, and dissemination
Ovarian carcinoma OVCAR-3VitroGel Hydrogel Matrix, VitroGel RGDCell proliferation, cell matrix interations
Ovarian OVCA429VitroGel Hydrogel Matrix, VitroGel COLMMP dependent cell invasion
Human osteosarcoma KHOSVitroGel Hydrogel Matrix, VitroGel 3Dcell proliferation and spheroids formation
Human osteosarcoma U2OSVitroGel Hydrogel Matrix, VitroGel 3Dcell proliferation and spheroids formation
Priess human lymphoblastoid cellsVitroGel Hydrogel Matrix, VitroGel 3DEnhance spheroids and cluster formation and promote cell viability.
CartilageChondrocytes (bovine)VitroGel Hydrogel Matrix, VitroGel RGDCell viability and proliferation
Chondrocytes (human)VitroGel Hydrogel Matrix, VitroGel RGDCell viability and proliferation
Connective tissueDermal fibroblasts (human)VitroGel Hydrogel Matrix, VitroGel RGDCell viability and spreading
VitroGel Hydrogel Matrix, VitroGel COLCell viability
Fibroblasts NIH3T3VitroGel Hydrogel Matrix, VitroGel RGDDirectional cell migration toward gradient
VitroGel Hydrogel Matrix, VitroGel COLCell spreading dependent on substrata rigidity
Foreskin fibroblasts (human)VitroGel Hydrogel Matrix, VitroGel RGDCell spreading
VitroGel Hydrogel Matrix, VitroGel YIGSRCell spreading
VitroGel Hydrogel Matrix, VitroGel MMPSubstrata degradation and cell invasion
Skin fibroblasts (skin)VitroGel Hydrogel Matrix, VitroGel IKVAVCell adhesion
Epidermal keratinocytesVitroGel Hydrogel Matrix, VitroGel COLCell viability
Epithelial CellsMouse ovarian follicle cellsVitroGel Hydrogel Matrix, VitroGel RGD3D cell culture using ES-hydrogel can enhance vitro follicle culture by considering the permeability and stiffness of the gel.
Human Nthy-ori 3-1 cellsVitroGel Hydrogel Matrix, VitroGel 3DEnhance spheroids and cluster formation and promote cell viability.
A549 cellsVitroGel Hydrogel Matrix, VitroGel RGDEnhance cell proliferation and cell matrix interactions.
MCF-12AVitroGel Hydrogel Matrix, VitroGel RGDEnhance cell proliferation and cell matrix interactions.
Immortalized bronchial epithelial cells HBEC-KRASVitroGel Hydrogel Matrix, VitroGel 3DCell proliferation
EyeCorneal endothelial B4G12VitroGel Hydrogel Matrix, VitroGel COLCell attachment and spreading
Retinal ganglion cells (xenopus)VitroGel Hydrogel Matrix, VitroGel COLNeurite outgrowth
Immune CellsCD8 + T cellsVitroGel Hydrogel Matrix, VitroGel 3DEnhance spheroids and cluster formation and promote cell viability.
KidneyHuman embryonic kidney HEK293VitroGel Hydrogel Matrix, VitroGel RGD3D spheroids formation
VitroGel Hydrogel Matrix, VitroGel COLCell proliferation and cluster growth
Madin-Darby Canine KidneyVitroGel Hydrogel Matrix, VitroGel RGDEpithelial cysts formation
podocytes (human)VitroGel Hydrogel Matrix, VitroGel COLGlomerular capillary formation
glomerular endothelial cells (human)VitroGel Hydrogel Matrix, VitroGel COLGlomerular capillary formation
LiverHepatocytes (human)VitroGel Hydrogel Matrix, VitroGel RGDFilopodia formation and synthesis of albumin
VitroGel Hydrogel Matrix, VitroGel COLCell attachment
Hepatocytes (mouse)VitroGel Hydrogel Matrix, VitroGel RGDCell viability
Hepatocytes (rat)VitroGel Hydrogel Matrix, VitroGel COLAlbumin secretion
Hepatocytes (swine)VitroGel Hydrogel Matrix, VitroGel COLCell spreading and albumin section
LungAlveolar basal epithelial A549VitroGel Hydrogel Matrix, VitroGel RGDCell attachment
Alveolar epithelial RLE-6TNVitroGel Hydrogel Matrix, VitroGel RGDCell attachment and mesenchymal differentiation
Pulmonary fibroblasts LL2VitroGel Hydrogel Matrix, VitroGel IKVAVCell adhesion
HFL1 lung fibroblasts CCL153VitroGel Hydrogel Matrix, VitroGel COLCell proliferation and spindle morphology
Lung cancer associated fibroblasts (human)VitroGel Hydrogel Matrix, VitroGel COLSubstrata contractility
Lung fibroblasts MCR-5VitroGel Hydrogel Matrix, VitroGel COLNGF-mediated substrata contraction
MuscleMyoblasts C2C12VitroGel Hydrogel Matrix, VitroGel RGDCell Proliferation and differentiation
VitroGel Hydrogel Matrix, VitroGel COLCell attachment, proliferation, and myofibril formation
VitroGel Hydrogel MatrixMyotube formation
VitroGel Hydrogel Matrix, VitroGel COLIntegrin dependent cell adhesion
Skeletal myoblasts (mouse)VitroGel Hydrogel Matrix, VitroGel RGDCell attachment, proliferation, and myofibril formation
Myoblasts (human)VitroGel Hydrogel Matrix, VitroGel COLCell adhesion, alignment along fiber, and myotube formation
Myoblasts C25Cl48VitroGel Hydrogel Matrix, VitroGel COLCell proliferation, differentiation and myotube formation
NeuralDorsal root ganglion (chick)VitroGel RGDNeurite formation and outgrowth
VitroGel COLForce dependent neurite outgrowth
Neural PC12VitroGel COLNeurite outgrowth
VitroGel IKVAVNeurite outgrowth
Neural stem cell/progenitor cell (rat)VitroGel YIGSRCell viability
VitroGel IKVAVCell attachment and differentiation
Neural stem cell/progenitor cell (human)VitroGel IKVAVCell viability and differentiation
VitroGel LDP1Cell viability and differentiation
VitroGel LDP1Cell viability
VitroGel COLCell attachment
Schwann cells (rat)VitroGel YIGSRCell attachment and migration
Neural stem cell/progenitor cell (mouse)VitroGel IKVAVCell adhesion and differentiation
Cortical astrocytes (rat)VitroGel IKVAVCell adhesion
Spiral ganglion neurons (mouse)VitroGel IKVAVNeurite outgrowth
Motor neurons (human)VitroGel COLForce dependent neurite outgrowth
Forebrain neurons (human)VitroGel COLForce dependent neurite outgrowth
Cortical neurons (rat)VitroGel COLNeuronal viability and neurite outgrowth
Dorsal root ganglion (rat)VitroGel COLNeurite outgrowth
Red Blood CellsRed Blood cellsVitroGel Hydrogel Matrix, VitroGel 3DEnhance spheroids and cluster formation and promote cell viability.
PancreasB-cells MIN6VitroGel Hydrogel Matrix, VitroGel IKVAVReduced apoptosis and increased insulin release
Stem cellsMesenchymal stem cells (human)VitroGel RGDCell viability
VitroGel RGDCell Proliferation and differentiation
VitroGel COLCell proliferation
VitroGel IKVAVNeuronal differentiation
VitroGel MMPNeuronal differentiation and neurite outgrowth
VitroGel COLCell attachment, spreading, viability, and osteoblast differentiation
Mesenchymal stem cells (mouse)VitroGel RGDCell spreading and migration
VitroGel MMPCell spreading and migration
Mesenchymal stem cells (rat)VitroGel RGDCell adhesion and spreading
Embryonic stem cells (mouse)VitroGel RGDEndothelial cell differentiation
VitroGel COLNeuronal differentiation and neurite outgrowth
VitroGel YIGSRNeuronal differentiation
Induced pluripotent stem cells (human)VitroGel YIGSRCell viability
VitroGel IKVAVCell viability
VitroGel LDP1Cell viability
Human IpscVitroGel RGDCell proliferation, and cell matrix interactions
Human stem cells from apical papilla SCAPVitroGel 3DCell viability
Adipose derived stem cells (human)VitroGel IKVAVCell attachment
Vascular/cardiacUmbilical vein endothelial cells (human)VitroGel RGDCell attachment, proliferation, migration, and angiogenesis
VitroGel YIGSRUpregulation in gene expression
VitroGel IKVAVMigratory cell infiltration
VitroGel MMPCell attachment, migration, and survival
VitroGel COLCell attachment, spreading, and VEGF dependent migration
Neonatal cardiac (rat)VitroGel RGDCell attachment and tissue regeneration
VitroGel YIGSRCell attachment similar to laminin
Aortic smooth muscle cells (human)VitroGel RGDCell attachment
Endothelial (human)VitroGel YIGSRCell differentiation
EndotheliocytesVitroGel YIGSRCell migration
Microvascular endothelial cells (human)VitroGel YIGSRCell mobility
Aortic endothelial cells (bovine)VitroGel COLForce dependent cell spreading
Capillary endothelial cells (bovine)VitroGel COLCapillary like network formation

Functional Hydrogel Types

References

[1]      A. Arjonen, R. Kaukonen, and J. Ivaska, “Filopodia and adhesion in cancer cell motility,” Cell Adhesion and Migration. 2011, doi: 10.4161/cam.5.5.17723.
[2]      M. Gimona, R. Buccione, S. A. Courtneidge, and S. Linder, “Assembly and biological role of podosomes and invadopodia,” Current Opinion in Cell Biology. 2008, doi: 10.1016/j.ceb.2008.01.005.
[3]      D. A. Murphy and S. A. Courtneidge, “The ‘ins’ and ‘outs’ of podosomes and invadopodia: Characteristics, formation and function,” Nature Reviews Molecular Cell Biology. 2011, doi: 10.1038/nrm3141.
[4]      C. M. Gould and S. A. Courtneidge, “Regulation of invadopodia by the tumor microenvironment,” Cell Adhesion and Migration. 2014, doi: 10.4161/cam.28346.
[5]      S. R. Caliari and J. A. Burdick, “A practical guide to hydrogels for cell culture,” Nature Methods. 2016, doi: 10.1038/nmeth.3839.
[6]      K. Wolf et al., “Collagen-based cell migration models in vitro and in vivo,” Seminars in Cell and Developmental Biology. 2009, doi: 10.1016/j.semcdb.2009.08.005.
[7]      S. L. Bellis, “Advantages of RGD peptides for directing cell association with biomaterials,” Biomaterials, 2011, doi: 10.1016/j.biomaterials.2011.02.029.
[8]      T. Maeda, K. Titani, and K. Sekiguchi, “Cell-adhesive activity and receptor-binding specificity of the laminin-derived YIGSR sequence grafted onto Staphylococcal protein a,” J. Biochem., 1994, doi: 10.1093/oxfordjournals.jbchem.a124315.
[9]      T. Y. Cheng, M. H. Chen, W. H. Chang, M. Y. Huang, and T. W. Wang, “Neural stem cells encapsulated in a functionalized self-assembling peptide hydrogel for brain tissue engineering,” Biomaterials, 2013, doi: 10.1016/j.biomaterials.2012.11.043.
[10]    J. Huang, “3D Cell Culture On VitroGel System,” Cytol. Tissue Biol., 2019, doi: 10.24966/ctb-9107/s1001.
[11]    K. Powell, “Adding depth to cell culture,” Science (80-. )., 2017, doi: 10.1126/science.opms.p1700113.
[12]    W. D. Mahauad-Fernandez and C. M. Okeoma, “B49, a BST-2-based peptide, inhibits adhesion and growth of breast cancer cells,” Sci. Rep., 2018, doi: 10.1038/s41598-018-22364-z.
[13]    P. Thanindratarn, X. Li, D. C. Dean, S. D. Nelson, F. J. Hornicek, and Z. Duan, “Establishment and Characterization of a Recurrent Osteosarcoma Cell Line: OSA 1777,” J. Orthop. Res., 2019, doi: 10.1002/jor.24528.
[14]    M. di Donato, G. Cernera, A. Migliaccio, and G. Castoria, “Nerve growth factor induces proliferation and aggressiveness in prostate cancer cells,” Cancers (Basel)., 2019, doi: 10.3390/cancers11060784.
[15]    E. J. Kim et al., “The new biocompatible material for mouse ovarian follicle development in three-dimensional in vitro culture systems,” Theriogenology, 2020, doi: 10.1016/j.theriogenology.2019.12.009.
[16]    Y. Yang et al., “Influence of Cell Spreading Area on the Osteogenic Commitment and Phenotype Maintenance of Mesenchymal Stem Cells,” Sci. Rep., 2019, doi: 10.1038/s41598-019-43362-9.
[17]    B. Shamloo et al., “Dysregulation of adenosine kinase isoforms in breast cancer,” Oncotarget, 2019, doi: 10.18632/oncotarget.27364.
[18]    F. Yang, C. G. Williams, D. A. Wang, H. Lee, P. N. Manson, and J. Elisseeff, “The effect of incorporating RGD adhesive peptide in polyethylene glycol diacrylate hydrogel on osteogenesis of bone marrow stromal cells,” Biomaterials, 2005, doi: 10.1016/j.biomaterials.2005.03.018.
[19]    D. Loessner, K. S. Stok, M. P. Lutolf, D. W. Hutmacher, J. A. Clements, and S. C. Rizzi, “Bioengineered 3D platform to explore cell-ECM interactions and drug resistance of epithelial ovarian cancer cells,” Biomaterials, 2010, doi: 10.1016/j.biomaterials.2010.07.064.
[20]    K. M. Park, Y. K. Joung, K. D. Park, S. Y. Lee, and M. C. Lee, “RGD-conjugated chitosan-pluronic hydrogels as a cell supported scaffold for articular cartilage regeneration,” Macromol. Res., 2008, doi: 10.1007/BF03218553.
[21]    X. Z. Shu et al., “Attachment and spreading of fibroblasts on an RGD peptide-modified injectable hyaluronan hydrogel,” J. Biomed. Mater. Res. – Part A, 2004, doi: 10.1002/jbm.a.20002.
[22]    E. Kim and W. B. Jeon, “Gene expression analysis of 3D spheroid culture of human embryonic kidney cells,” Toxicol. Environ. Health Sci., 2013, doi: 10.1007/s13530-013-0160-y.
[23]    L. De Bartolo et al., “Biotransformation and liver-specific functions of human hepatocytes in culture on RGD-immobilized plasma-processed membranes,” Biomaterials, 2005, doi: 10.1016/j.biomaterials.2004.11.009.
[24]    J. A. Rowley, G. Madlambayan, and D. J. Mooney, “Alginate hydrogels as synthetic extracellular matrix materials,” Biomaterials, 1999, doi: 10.1016/S0142-9612(98)00107-0.
[25]    K. J. Lampe, A. L. Antaris, and S. C. Heilshorn, “Design of three-dimensional engineered protein hydrogels for tailored control of neurite growth,” Acta Biomater., 2013, doi: 10.1016/j.actbio.2012.10.033.
[26]    L. Schukur, P. Zorlutuna, J. M. Cha, H. Bae, and A. Khademhosseini, “Directed Differentiation of Size-Controlled Embryoid Bodies Towards Endothelial and Cardiac Lineages in RGD-Modified Poly(Ethylene Glycol) Hydrogels,” Adv. Healthc. Mater., 2013, doi: 10.1002/adhm.201200194.
[27]    N. Zhao, M. R. Battig, M. Xu, X. Wang, N. Xiong, and Y. Wang, “Development of a Dual-Functional Hydrogel Using RGD and Anti-VEGF Aptamer,” Macromol. Biosci., 2017, doi: 10.1002/mabi.201700201.
[28]    N. Davidenko et al., “Selecting the correct cellular model for assessing of the biological response of collagen-based biomaterials,” Acta Biomater., 2018, doi: 10.1016/j.actbio.2017.10.035.
[29]    J. D. Malcor et al., “The synthesis and coupling of photoreactive collagen-based peptides to restore integrin reactivity to an inert substrate, chemically-crosslinked collagen,” Biomaterials, 2016, doi: 10.1016/j.biomaterials.2016.01.044.
[30]    J. T. Connelly, T. A. Petrie, A. J. García, and M. E. Levenston, “Fibronectin-and collagen-mimetic ligands regulate bone marrow stromal cell chondrogenesis in three-dimensional hydrogels,” Eur. Cells Mater., 2011, doi: 10.22203/ecm.v022a13.
[31]    C. D. Reyes, T. A. Petrie, K. L. Burns, Z. Schwartz, and A. J. García, “Biomolecular surface coating to enhance orthopaedic tissue healing and integration,” Biomaterials, 2007, doi: 10.1016/j.biomaterials.2007.04.003.
[32]    M. S. Rehmann, J. I. Luna, E. Maverakis, and A. M. Kloxin, “Tuning microenvironment modulus and biochemical composition promotes human mesenchymal stem cell tenogenic differentiation,” J. Biomed. Mater. Res. – Part A, 2016, doi: 10.1002/jbm.a.35650.
[33]    A. Shekaran et al., “Bone regeneration using an alpha 2 beta 1 integrin-specific hydrogel as a BMP-2 delivery vehicle,” Biomaterials, 2014, doi: 10.1016/j.biomaterials.2014.03.055.
[34]    L. A. Sawicki et al., “Tunable synthetic extracellular matrices to investigate breast cancer response to biophysical and biochemical cues,” APL Bioeng., 2019, doi: 10.1063/1.5064596.
[35]    A. V. Taubenberger et al., “3D extracellular matrix interactions modulate tumour cell growth, invasion and angiogenesis in engineered tumour microenvironments,” Acta Biomater., 2016, doi: 10.1016/j.actbio.2016.03.017.
[36]    L. Kam, W. Shain, J. N. Turner, and R. Bizios, “Selective adhesion of astrocytes to surfaces modified with immobilized peptides,” Biomaterials, 2002, doi: 10.1016/S0142-9612(01)00133-8.
[37]    M. R. Hynd, J. P. Frampton, N. Dowell-Mesfin, J. N. Turner, and W. Shain, “Directed cell growth on protein-functionalized hydrogel surfaces,” J. Neurosci. Methods, 2007, doi: 10.1016/j.jneumeth.2007.01.024.
[38]    A. Farrukh et al., “Bifunctional Hydrogels Containing the Laminin Motif IKVAV Promote Neurogenesis,” Stem Cell Reports, 2017, doi: 10.1016/j.stemcr.2017.09.002.
[39]    L. Y. Santiago, R. W. Nowak, J. P. Rubin, and K. G. Marra, “Peptide-surface modification of poly(caprolactone) with laminin-derived sequences for adipose-derived stem cell applications,” Biomaterials, 2006, doi: 10.1016/j.biomaterials.2006.01.011.
[40]    E. M. do. Reis, F. V. Berti, G. Colla, and L. M. Porto, “Bacterial nanocellulose-IKVAV hydrogel matrix modulates melanoma tumor cell adhesion and proliferation and induces vasculogenic mimicry in vitro,” J. Biomed. Mater. Res. – Part B Appl. Biomater., 2018, doi: 10.1002/jbm.b.34055.
[41]    W. Sun, T. Incitti, C. Migliaresi, A. Quattrone, S. Casarosa, and A. Motta, “Viability and neuronal differentiation of neural stem cells encapsulated in silk fibroin hydrogel functionalized with an IKVAV peptide,” J. Tissue Eng. Regen. Med., 2017, doi: 10.1002/term.2053.
[42]    R. Bellamkonda, J. P. Ranieri, and P. Aebischer, “Laminin oligopeptide derivatized agarose gels allow three‐dimensional neurite extension in vitro,” J. Neurosci. Res., 1995, doi: 10.1002/jnr.490410409.
[43]    C. Frick et al., “Biofunctionalized peptide-based hydrogels provide permissive scaffolds to attract neurite outgrowth from spiral ganglion neurons,” Colloids Surfaces B Biointerfaces, 2017, doi: 10.1016/j.colsurfb.2016.10.003.
[44]    W. Sun et al., “Co-culture of outgrowth endothelial cells with human mesenchymal stem cells in silk fibroin hydrogels promotes angiogenesis,” Biomed. Mater., 2016, doi: 10.1088/1748-6041/11/3/035009.
[45]    Y. Iwamoto et al., “YIGSR, a synthetic laminin pentapeptide, inhibits experimental metastasis formation,” Science (80-. )., 1987, doi: 10.1126/science.2961059.
[46]    M. Nomizu, K. Yamamura, H. K. Kleinman, and Y. Yamada, “Multimeric Forms of Tyr-Ile-Gly-Ser-Arg (YIGSR) Peptide Enhance the Inhibition of Tumor Growth and Metastasis,” Cancer Res., 1993.
[47]    N. Yoshida et al., “The laminin-derived peptide YIGSR (Tyr-Ile-Gly-Ser-Arg) inhibits human pre-B leukaemic cell growth and dissemination to organs in SCID mice,” Br. J. Cancer, 1999, doi: 10.1038/sj.bjc.6690618.
[48]    S. P. Massia, S. S. Rao, and J. A. Hubbell, “Covalently immobilized laminin peptide Tyr-Ile-Gly-Ser-Arg (YIGSR) supports cell spreading and co-localization of the 67-kilodalton laminin receptor with α-actinin and vinculin,” J. Biol. Chem., 1993.
[49]    C. M. M. Motta, K. J. Endres, C. Wesdemiotis, R. K. Willits, and M. L. Becker, “Enhancing Schwann cell migration using concentration gradients of laminin-derived peptides,” Biomaterials, 2019, doi: 10.1016/j.biomaterials.2019.119335.
[50]    S. Y. Boateng, S. S. Lateef, W. Mosley, T. J. Hartman, L. Hanley, and B. Russell, “RGD and YIGSR synthetic peptides facilitate cellular adhesion identical to that of laminin and fibronectin but alter the physiology of neonatal cardiac myocytes,” Am. J. Physiol. – Cell Physiol., 2005, doi: 10.1152/ajpcell.00199.2004.
[51]    Y. KUBOTA and M. MIZOGUCHI, “Modulation of morphological differentiation of human endothelial cells in culture by the synthetic peptide YIGSR and cytochalasin B,” Clin. Exp. Dermatol., 1993, doi: 10.1111/j.1365-2230.1993.tb02177.x.
[52]    T. Ren, S. Yu, Z. Mao, S. E. Moya, L. Han, and C. Gao, “Complementary density gradient of poly(hydroxyethyl methacrylate) and YIGSR selectively guides migration of endotheliocytes,” Biomacromolecules, 2014, doi: 10.1021/bm500385n.
[53]    J. Lam, S. T. Carmichael, W. E. Lowry, and T. Segura, “Hydrogel design of experiments methodology to optimize hydrogel for iPSC-NPC culture,” Adv. Healthc. Mater., 2015, doi: 10.1002/adhm.201400410.
[54]    M. H. Fittkau et al., “The selective modulation of endothelial cell mobility on RGD peptide containing surfaces by YIGSR peptides,” Biomaterials, 2005, doi: 10.1016/j.biomaterials.2004.02.012.
[55]    C. C. Horgan et al., “Characterisation of minimalist co-assembled fluorenylmethyloxycarbonyl self-assembling peptide systems for presentation of multiple bioactive peptides,” Acta Biomater., 2016, doi: 10.1016/j.actbio.2016.04.038.
[56]    L. Romagnano and B. Babiarz, “Mechanisms of Murine Trophoblast Interaction with Laminin1,” Biol. Reprod., 1993, doi: 10.1095/biolreprod49.2.374.
[57]    P. Lu, K. Takai, V. M. Weaver, and Z. Werb, “Extracellular Matrix degradation and remodeling in development and disease,” Cold Spring Harb. Perspect. Biol., 2011, doi: 10.1101/cshperspect.a005058.
[58]    K. M. Park, D. Lewis, and S. Gerecht, “Bioinspired Hydrogels to Engineer Cancer Microenvironments,” Annu. Rev. Biomed. Eng., 2017, doi: 10.1146/annurev-bioeng-071516-044619.
[59]    Y. Lei, S. Gojgini, J. Lam, and T. Segura, “The spreading, migration and proliferation of mouse mesenchymal stem cells cultured inside hyaluronic acid hydrogels,” Biomaterials, 2011, doi: 10.1016/j.biomaterials.2010.08.103.
[60]    T. P. Kraehenbuehl, L. S. Ferreira, P. Zammaretti, J. A. Hubbell, and R. Langer, “Cell-responsive hydrogel for encapsulation of vascular cells,” Biomaterials, 2009, doi: 10.1016/j.biomaterials.2009.04.057.
[61]    M. P. Lutolf et al., “Synthetic matrix metalloproteinase-sensitive hydrogels for the conduction of tissue regeneration: Engineering cell-invasion characteristics,” Proc. Natl. Acad. Sci. U. S. A., 2003, doi: 10.1073/pnas.0737381100.
[62]    S. A. Fisher, P. N. Anandakumaran, S. C. Owen, and M. S. Shoichet, “Tuning the Microenvironment: Click-Crosslinked Hyaluronic Acid-Based Hydrogels Provide a Platform for Studying Breast Cancer Cell Invasion,” Adv. Funct. Mater., 2015, doi: 10.1002/adfm.201502778.
[63]    S. P. Singh et al., “A synthetic modular approach for modeling the role of the 3D microenvironment in tumor progression,” Sci. Rep., 2015, doi: 10.1038/srep17814.
[64]    S. B. Anderson, C. C. Lin, D. V. Kuntzler, and K. S. Anseth, “The performance of human mesenchymal stem cells encapsulated in cell-degradable polymer-peptide hydrogels,” Biomaterials, 2011, doi: 10.1016/j.biomaterials.2011.01.064.
[65]    S. Khetan, M. Guvendiren, W. R. Legant, D. M. Cohen, C. S. Chen, and J. A. Burdick, “Degradation-mediated cellular traction directs stem cell fate in covalently crosslinked three-dimensional hydrogels,” Nat. Mater., 2013, doi: 10.1038/nmat3586.
[66]    T. Rozario and D. W. DeSimone, “The extracellular matrix in development and morphogenesis: A dynamic view,” Developmental Biology. 2010, doi: 10.1016/j.ydbio.2009.10.026.
[67]    C. Bonnans, J. Chou, and Z. Werb, “Remodelling the extracellular matrix in development and disease,” Nature Reviews Molecular Cell Biology. 2014, doi: 10.1038/nrm3904.
[68]    N. V. Krakhmal, M. V. Zavyalova, E. V. Denisov, S. V. Vtorushin, and V. M. Perelmuter, “Cancer invasion: Patterns and mechanisms,” Acta Naturae. 2015, doi: 10.32607/20758251-2015-7-2-17-28.