3D Cell Culture

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Tsuyoshi Takato - One of the best experts on this subject based on the ideXlab platform.

  • sugar based crosslinker forms a stable atelocollagen hydrogel that is a favorable microenvironment for 3D Cell Culture
    Journal of Biomedical Materials Research Part A, 2014
    Co-Authors: Wataru Kamimura, Hiroyuki Koyama, Tetsuro Miyata, Tsuyoshi Takato
    Abstract:

    We created sugar-based crosslinkers for precise chemical crosslinking of atelocollagen to form a stable hydrogel microenvironment for three-dimensional (3D) Cell Culture. Crosslinkers were synthesized by partially oxidizing glucose, fructose, maltose, sucrose, or raffinose with periodate. The partial oxidization of sugar generated multiple aldehydes in the molecule, which acted as multifunctional crosslinkers, allowing atelocollagen to form chemical hydrogels. The crosslink reaction of atelocollagen was competitively suppressed by the addition of amino acid-rich medium, enabling flexible control of 3D molecular density in the acquired hydrogel. To evaluate structural stability, ultrasonic stress was loaded onto the acquired hydrogel and sequential measurement of water content showed that the crosslinking considerably stabilized the hydrogel structure. The effectiveness of the atelocollagen hydrogel as a 3D Culture scaffold was analyzed by embedding and culturing endothelial Cells or cardiomyocytes in it. Endothelial Cells formed 3D capillary-like structures at 12 h, and cardiomyocytes formed a beating 3D netlike structure by 7 days. These findings suggest that crosslinked atelocollagen hydrogel effectively functions as a stable scaffold in 3D Culture. © 2014 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 102A: 4309–4316, 2014.

  • Sugar‐based crosslinker forms a stable atelocollagen hydrogel that is a favorable microenvironment for 3D Cell Culture
    Journal of Biomedical Materials Research Part A, 2014
    Co-Authors: Wataru Kamimura, Hiroyuki Koyama, Tetsuro Miyata, Tsuyoshi Takato
    Abstract:

    We created sugar-based crosslinkers for precise chemical crosslinking of atelocollagen to form a stable hydrogel microenvironment for three-dimensional (3D) Cell Culture. Crosslinkers were synthesized by partially oxidizing glucose, fructose, maltose, sucrose, or raffinose with periodate. The partial oxidization of sugar generated multiple aldehydes in the molecule, which acted as multifunctional crosslinkers, allowing atelocollagen to form chemical hydrogels. The crosslink reaction of atelocollagen was competitively suppressed by the addition of amino acid-rich medium, enabling flexible control of 3D molecular density in the acquired hydrogel. To evaluate structural stability, ultrasonic stress was loaded onto the acquired hydrogel and sequential measurement of water content showed that the crosslinking considerably stabilized the hydrogel structure. The effectiveness of the atelocollagen hydrogel as a 3D Culture scaffold was analyzed by embedding and culturing endothelial Cells or cardiomyocytes in it. Endothelial Cells formed 3D capillary-like structures at 12 h, and cardiomyocytes formed a beating 3D netlike structure by 7 days. These findings suggest that crosslinked atelocollagen hydrogel effectively functions as a stable scaffold in 3D Culture. © 2014 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 102A: 4309–4316, 2014.

Wataru Kamimura - One of the best experts on this subject based on the ideXlab platform.

  • sugar based crosslinker forms a stable atelocollagen hydrogel that is a favorable microenvironment for 3D Cell Culture
    Journal of Biomedical Materials Research Part A, 2014
    Co-Authors: Wataru Kamimura, Hiroyuki Koyama, Tetsuro Miyata, Tsuyoshi Takato
    Abstract:

    We created sugar-based crosslinkers for precise chemical crosslinking of atelocollagen to form a stable hydrogel microenvironment for three-dimensional (3D) Cell Culture. Crosslinkers were synthesized by partially oxidizing glucose, fructose, maltose, sucrose, or raffinose with periodate. The partial oxidization of sugar generated multiple aldehydes in the molecule, which acted as multifunctional crosslinkers, allowing atelocollagen to form chemical hydrogels. The crosslink reaction of atelocollagen was competitively suppressed by the addition of amino acid-rich medium, enabling flexible control of 3D molecular density in the acquired hydrogel. To evaluate structural stability, ultrasonic stress was loaded onto the acquired hydrogel and sequential measurement of water content showed that the crosslinking considerably stabilized the hydrogel structure. The effectiveness of the atelocollagen hydrogel as a 3D Culture scaffold was analyzed by embedding and culturing endothelial Cells or cardiomyocytes in it. Endothelial Cells formed 3D capillary-like structures at 12 h, and cardiomyocytes formed a beating 3D netlike structure by 7 days. These findings suggest that crosslinked atelocollagen hydrogel effectively functions as a stable scaffold in 3D Culture. © 2014 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 102A: 4309–4316, 2014.

  • Sugar‐based crosslinker forms a stable atelocollagen hydrogel that is a favorable microenvironment for 3D Cell Culture
    Journal of Biomedical Materials Research Part A, 2014
    Co-Authors: Wataru Kamimura, Hiroyuki Koyama, Tetsuro Miyata, Tsuyoshi Takato
    Abstract:

    We created sugar-based crosslinkers for precise chemical crosslinking of atelocollagen to form a stable hydrogel microenvironment for three-dimensional (3D) Cell Culture. Crosslinkers were synthesized by partially oxidizing glucose, fructose, maltose, sucrose, or raffinose with periodate. The partial oxidization of sugar generated multiple aldehydes in the molecule, which acted as multifunctional crosslinkers, allowing atelocollagen to form chemical hydrogels. The crosslink reaction of atelocollagen was competitively suppressed by the addition of amino acid-rich medium, enabling flexible control of 3D molecular density in the acquired hydrogel. To evaluate structural stability, ultrasonic stress was loaded onto the acquired hydrogel and sequential measurement of water content showed that the crosslinking considerably stabilized the hydrogel structure. The effectiveness of the atelocollagen hydrogel as a 3D Culture scaffold was analyzed by embedding and culturing endothelial Cells or cardiomyocytes in it. Endothelial Cells formed 3D capillary-like structures at 12 h, and cardiomyocytes formed a beating 3D netlike structure by 7 days. These findings suggest that crosslinked atelocollagen hydrogel effectively functions as a stable scaffold in 3D Culture. © 2014 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 102A: 4309–4316, 2014.

Ovijit Chaudhuri - One of the best experts on this subject based on the ideXlab platform.

  • varying peg density to control stress relaxation in alginate peg hydrogels for 3D Cell Culture studies
    Biomaterials, 2019
    Co-Authors: Sungmin Nam, Ryan S Stowers, Junzhe Lou, Yan Xia, Ovijit Chaudhuri
    Abstract:

    Hydrogels are commonly used as artificial extraCellular matrices for 3D Cell Culture and for tissue engineering. Viscoelastic hydrogels with tunable stress relaxation have recently been developed, and stress relaxation in the hydrogels has been found to play a key role in regulating Cell behaviors such as differentiation, spreading, and proliferation. Here we report a simple but precise materials approach to tuning stress relaxation of alginate hydrogels with polyethylene glycol (PEG) covalently grafted onto the alginate. Hydrogel relaxation was modulated independent of the initial elastic modulus by varying molecular weight and concentration of PEG along with calcium crosslinking of the alginate. Increased concentration and molecular weight of the PEG resulted in faster stress relaxation, a higher loss modulus, and increased creep. Interestingly, we found that stress relaxation of the hydrogels is determined by the total mass amount of PEG in the hydrogel, and not the molecular weight or concentration of PEG chains alone. We then evaluated the utility of these hydrogels for 3D Cell Culture. Faster relaxation in RGD-coupled alginate-PEG hydrogels led to increased spreading and proliferation of fibroblasts, and enhanced osteogenic differentiation of mesenchymal stem Cells (MSCs). Thus, this work establishes a new materials approach to tuning stress relaxation in alginate hydrogels for 3D Cell Culture.

  • stress relaxing hyaluronic acid collagen hydrogels promote Cell spreading fiber remodeling and focal adhesion formation in 3D Cell Culture
    Biomaterials, 2018
    Co-Authors: Junzhe Lou, Ryan S Stowers, Sungmin Nam, Yan Xia, Ovijit Chaudhuri
    Abstract:

    The physical and architectural cues of the extraCellular matrix (ECM) play a critical role in regulating important Cellular functions such as spreading, migration, proliferation, and differentiation. Natural ECM is a complex viscoelastic scaffold composed of various distinct components that are often organized into a fibrillar microstructure. Hydrogels are frequently used as synthetic ECMs for 3D Cell Culture, but are typically elastic, due to covalent crosslinking, and non-fibrillar. Recent work has revealed the importance of stress relaxation in viscoelastic hydrogels in regulating biological processes such as spreading and differentiation, but these studies all utilize synthetic ECM hydrogels that are non-fibrillar. Key mechanotransduction events, such as focal adhesion formation, have only been observed in fibrillar networks in 3D Culture to date. Here we present an interpenetrating network (IPN) hydrogel system based on HA crosslinked with dynamic covalent bonds and collagen I that captures the viscoelasticity and fibrillarity of ECM in tissues. The IPN hydrogels exhibit two distinct processes in stress relaxation, one from collagen and the other from HA crosslinking dynamics. Stress relaxation in the IPN hydrogels can be tuned by modulating HA crosslinker affinity, molecular weight of the HA, or HA concentration. Faster relaxation in the IPN hydrogels promotes Cell spreading, fiber remodeling, and focal adhesion (FA) formation - behaviors often inhibited in other hydrogel-based materials in 3D Culture. This study presents a new, broadly adaptable materials platform for mimicking key ECM features of viscoelasticity and fibrillarity in hydrogels for 3D Cell Culture and sheds light on how these mechanical and structural cues regulate Cell behavior.

  • viscoelastic hydrogels for 3D Cell Culture
    Biomaterials Science, 2017
    Co-Authors: Ovijit Chaudhuri
    Abstract:

    In tissues, many Cells are surrounded by and interact with a three-dimensional soft extraCellular matrix (ECM). Both the physical and biochemical properties of the ECM play a major role in regulating Cell behaviours. To better understand the impact of ECM properties on Cell behaviours, natural and synthetic hydrogels have been developed for use as synthetic ECMs for 3D Cell Culture. It has long been known that ECM and tissues are viscoelastic, or display a time-dependent response to deformation or mechanical loading, exhibiting stress relaxation and creep. However, only recently have there been efforts made to understand the role of the time-dependent aspects of the ECM mechanics on regulating Cell behaviours using hydrogels for 3D Culture. Here we review the characterization and molecular basis of hydrogel viscoelasticity and plasticity, and describe newly developed approaches to tuning viscoelasticity in hydrogels for 2D and 3D Culture. Then we highlight several recent studies finding a potent impact of hydrogel stress relaxation or creep on Cell behaviours such as Cell spreading, proliferation, and differentiation of mesenchymal stem Cells. The role of time-dependent mechanics on Cell biology remains largely unclear, and ripe for further exploration. Further elucidation of this topic may substantially advance our understanding of Cell-matrix interactions during development, homeostasis, wound healing, and disease, and guide the design of biomaterials for regenerative medicine.

  • 3D Cell Culture in Interpenetrating Networks of Alginate and rBM Matrix.
    Methods in molecular biology (Clifton N.J.), 2017
    Co-Authors: Katrina M. Wisdom, Ovijit Chaudhuri
    Abstract:

    Altered tissue mechanical properties have been implicated in many key physiological and pathological processes. Hydrogel-based materials systems, made with native extraCellular matrix (ECM) proteins, nonnative biopolymers, or synthetic polymers are often used to study these processes in vitro in 3D Cell Culture experiments. However, each of these materials systems present major limitations when used in mechanobiological studies. While native ECM-based hydrogels may enable good recapitulation of physiological behavior, the mechanics of these hydrogels are often manipulated by increasing or decreasing the protein concentration. This manipulation changes Cell adhesion ligand density, thereby altering Cell signaling. Alternatively, synthetic polymer-based hydrogels and nonnative biopolymer-based hydrogels can be mechanically tuned and engineered to present Cell adhesion peptide motifs, but still may not fully promote physiologically relevant behavior. Here, we combine the advantages of native ECM proteins and nonnative biopolymers in interpenetrating network (IPN) hydrogels consisting of rBM matrix, which contains ligands native to epithelial basement membrane, and alginate, an inert biopolymer derived from seaweed. The following protocol details the generation of IPNs for mechanical testing or for 3D Cell Culture. This biomaterial system offers the ability to tune the stiffness of the 3D microenvironment without altering Cell adhesion ligand concentration or pore size.

Hiroyuki Koyama - One of the best experts on this subject based on the ideXlab platform.

  • sugar based crosslinker forms a stable atelocollagen hydrogel that is a favorable microenvironment for 3D Cell Culture
    Journal of Biomedical Materials Research Part A, 2014
    Co-Authors: Wataru Kamimura, Hiroyuki Koyama, Tetsuro Miyata, Tsuyoshi Takato
    Abstract:

    We created sugar-based crosslinkers for precise chemical crosslinking of atelocollagen to form a stable hydrogel microenvironment for three-dimensional (3D) Cell Culture. Crosslinkers were synthesized by partially oxidizing glucose, fructose, maltose, sucrose, or raffinose with periodate. The partial oxidization of sugar generated multiple aldehydes in the molecule, which acted as multifunctional crosslinkers, allowing atelocollagen to form chemical hydrogels. The crosslink reaction of atelocollagen was competitively suppressed by the addition of amino acid-rich medium, enabling flexible control of 3D molecular density in the acquired hydrogel. To evaluate structural stability, ultrasonic stress was loaded onto the acquired hydrogel and sequential measurement of water content showed that the crosslinking considerably stabilized the hydrogel structure. The effectiveness of the atelocollagen hydrogel as a 3D Culture scaffold was analyzed by embedding and culturing endothelial Cells or cardiomyocytes in it. Endothelial Cells formed 3D capillary-like structures at 12 h, and cardiomyocytes formed a beating 3D netlike structure by 7 days. These findings suggest that crosslinked atelocollagen hydrogel effectively functions as a stable scaffold in 3D Culture. © 2014 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 102A: 4309–4316, 2014.

  • Sugar‐based crosslinker forms a stable atelocollagen hydrogel that is a favorable microenvironment for 3D Cell Culture
    Journal of Biomedical Materials Research Part A, 2014
    Co-Authors: Wataru Kamimura, Hiroyuki Koyama, Tetsuro Miyata, Tsuyoshi Takato
    Abstract:

    We created sugar-based crosslinkers for precise chemical crosslinking of atelocollagen to form a stable hydrogel microenvironment for three-dimensional (3D) Cell Culture. Crosslinkers were synthesized by partially oxidizing glucose, fructose, maltose, sucrose, or raffinose with periodate. The partial oxidization of sugar generated multiple aldehydes in the molecule, which acted as multifunctional crosslinkers, allowing atelocollagen to form chemical hydrogels. The crosslink reaction of atelocollagen was competitively suppressed by the addition of amino acid-rich medium, enabling flexible control of 3D molecular density in the acquired hydrogel. To evaluate structural stability, ultrasonic stress was loaded onto the acquired hydrogel and sequential measurement of water content showed that the crosslinking considerably stabilized the hydrogel structure. The effectiveness of the atelocollagen hydrogel as a 3D Culture scaffold was analyzed by embedding and culturing endothelial Cells or cardiomyocytes in it. Endothelial Cells formed 3D capillary-like structures at 12 h, and cardiomyocytes formed a beating 3D netlike structure by 7 days. These findings suggest that crosslinked atelocollagen hydrogel effectively functions as a stable scaffold in 3D Culture. © 2014 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 102A: 4309–4316, 2014.

Tetsuro Miyata - One of the best experts on this subject based on the ideXlab platform.

  • sugar based crosslinker forms a stable atelocollagen hydrogel that is a favorable microenvironment for 3D Cell Culture
    Journal of Biomedical Materials Research Part A, 2014
    Co-Authors: Wataru Kamimura, Hiroyuki Koyama, Tetsuro Miyata, Tsuyoshi Takato
    Abstract:

    We created sugar-based crosslinkers for precise chemical crosslinking of atelocollagen to form a stable hydrogel microenvironment for three-dimensional (3D) Cell Culture. Crosslinkers were synthesized by partially oxidizing glucose, fructose, maltose, sucrose, or raffinose with periodate. The partial oxidization of sugar generated multiple aldehydes in the molecule, which acted as multifunctional crosslinkers, allowing atelocollagen to form chemical hydrogels. The crosslink reaction of atelocollagen was competitively suppressed by the addition of amino acid-rich medium, enabling flexible control of 3D molecular density in the acquired hydrogel. To evaluate structural stability, ultrasonic stress was loaded onto the acquired hydrogel and sequential measurement of water content showed that the crosslinking considerably stabilized the hydrogel structure. The effectiveness of the atelocollagen hydrogel as a 3D Culture scaffold was analyzed by embedding and culturing endothelial Cells or cardiomyocytes in it. Endothelial Cells formed 3D capillary-like structures at 12 h, and cardiomyocytes formed a beating 3D netlike structure by 7 days. These findings suggest that crosslinked atelocollagen hydrogel effectively functions as a stable scaffold in 3D Culture. © 2014 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 102A: 4309–4316, 2014.

  • Sugar‐based crosslinker forms a stable atelocollagen hydrogel that is a favorable microenvironment for 3D Cell Culture
    Journal of Biomedical Materials Research Part A, 2014
    Co-Authors: Wataru Kamimura, Hiroyuki Koyama, Tetsuro Miyata, Tsuyoshi Takato
    Abstract:

    We created sugar-based crosslinkers for precise chemical crosslinking of atelocollagen to form a stable hydrogel microenvironment for three-dimensional (3D) Cell Culture. Crosslinkers were synthesized by partially oxidizing glucose, fructose, maltose, sucrose, or raffinose with periodate. The partial oxidization of sugar generated multiple aldehydes in the molecule, which acted as multifunctional crosslinkers, allowing atelocollagen to form chemical hydrogels. The crosslink reaction of atelocollagen was competitively suppressed by the addition of amino acid-rich medium, enabling flexible control of 3D molecular density in the acquired hydrogel. To evaluate structural stability, ultrasonic stress was loaded onto the acquired hydrogel and sequential measurement of water content showed that the crosslinking considerably stabilized the hydrogel structure. The effectiveness of the atelocollagen hydrogel as a 3D Culture scaffold was analyzed by embedding and culturing endothelial Cells or cardiomyocytes in it. Endothelial Cells formed 3D capillary-like structures at 12 h, and cardiomyocytes formed a beating 3D netlike structure by 7 days. These findings suggest that crosslinked atelocollagen hydrogel effectively functions as a stable scaffold in 3D Culture. © 2014 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 102A: 4309–4316, 2014.