Photopatterning of hydrogel scaffolds coupled to filter materials using stereolithography for perfused 3D culture of hepatocytes

Abstract The development of livers-on-a-chip aims to provide pharmaceutical companies with reliable systems to perform drug screening and toxicological studies. To that end, microfluidic systems are engineered to mimic the functions and architecture of this organ. In this context we have designed a device that reproduces series of liver microarchitectures, each permitting the 3D culture of hepatocytes by confining them to a chamber that is separated from the medium conveying channel by very thin slits. We modified the structure to ensure its compatibility with the culture of hepatocytes from different sources. Our device was adapted to the migratory and adhesion properties of HepaRG cells at various stages of differentiation. To prevent hepatoblast-like cells from migrating out of the chambers, the slit height was decreased and the medium flow rate increased. Maintaining already differentiated and less adherent cells within the chambers required desensitisation of the system to pressure variations. Using this device, it was possible to keep the cells alive for more than 14 days, during which they achieved a 3D organisation and acquired or maintained their differentiation into hepatocytes. With its multiple micro-chambers for hepatocyte culture, this microfluidic device architecture offers a promising opportunity to provide new tools for drug screening applications.

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Use of hydrogel scaffolds to develop an in vitro 3D culture model of human intestinal epithelium

Acta Biomaterialia ◽

10.1016/j.actbio.2017.08.035 ◽

2017 ◽

Vol 62 ◽

pp. 128-143 ◽

Cited By ~ 18

Author(s):

R.H. Dosh ◽

A. Essa ◽

N. Jordan-Mahy ◽

C. Sammon ◽

C.L. Le Maitre

Keyword(s):

Intestinal Epithelium ◽

3D Culture ◽

Hydrogel Scaffolds ◽

Culture Model ◽

3D Culture Model ◽

Human Intestinal Epithelium

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Reconstruction of Liver Organoid Using an Apatite-Fiber Scaffold, a Radial-Flow Bioreactor and FCL-4 Cells of Hepatocyte Model

Key Engineering Materials ◽

10.4028/www.scientific.net/kem.361-363.1165 ◽

2007 ◽

Vol 361-363 ◽

pp. 1165-1168 ◽

Cited By ~ 1

Author(s):

Mamoru Aizawa ◽

A. Hiramoto ◽

H. Maehashi ◽

Tomokazu Matsuura

Keyword(s):

Tissue Engineering ◽

Single Crystal ◽

Bone Tissue Engineering ◽

Radial Flow ◽

Three Dimensional ◽

3D Culture ◽

High Porosity ◽

Fiber Scaffolds ◽

Radial Flow Bioreactor ◽

Culture Of Hepatocytes

We have previously developed apatite-fiber scaffolds (AFSs) for bone tissue engineering using single-crystal apatite fibers and carbon beads. In the present investigation, we examined the possibility of reconstruction of a liver organoid via three-dimensional (3D) culture of hepatocytes using the AFSs and the radial-flow bioreactor (RFB), aiming to apply the scaffold as a matrix for regeneration of a real organ. FLC-4 cells were used as a model of hepatocyte. The cells were well-viable in the RFB settled with AFSs over a period for 28 d, compared with the cases of cellulose beads and apatite beads with high porosity of 85%. We conclude that the present AFS may be a promising scaffold for tissue engineering of liver.

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Bioprintable Lung Extracellular Matrix Hydrogel Scaffolds for 3D Culture of Mesenchymal Stromal Cells

Polymers ◽

10.3390/polym13142350 ◽

2021 ◽

Vol 13 (14) ◽

pp. 2350

Author(s):

Bryan Falcones ◽

Héctor Sanz-Fraile ◽

Esther Marhuenda ◽

Irene Mendizábal ◽

Ignacio Cabrera-Aguilera ◽

...

Keyword(s):

Extracellular Matrix ◽

Cultured Cells ◽

3D Culture ◽

Focal Adhesions ◽

Fold Increase ◽

Paracrine Factors ◽

Hydrogel Scaffolds ◽

Vitro Model ◽

Cells Cultured

Mesenchymal stromal cell (MSC)-based cell therapy in acute respiratory diseases is based on MSC secretion of paracrine factors. Several strategies have proposed to improve this are being explored including pre-conditioning the MSCs prior to administration. We here propose a strategy for improving the therapeutic efficacy of MSCs based on cell preconditioning by growing them in native extracellular matrix (ECM) derived from the lung. To this end, a bioink with tunable stiffness based on decellularized porcine lung ECM hydrogels was developed and characterized. The bioink was suitable for 3D culturing of lung-resident MSCs without the need for additional chemical or physical crosslinking. MSCs showed good viability, and contraction assays showed the existence of cell–matrix interactions in the bioprinted scaffolds. Adhesion capacity and length of the focal adhesions formed were increased for the cells cultured within the lung hydrogel scaffolds. Also, there was more than a 20-fold increase of the expression of the CXCR4 receptor in the 3D-cultured cells compared to the cells cultured in plastic. Secretion of cytokines when cultured in an in vitro model of lung injury showed a decreased secretion of pro-inflammatory mediators for the cells cultured in the 3D scaffolds. Moreover, the morphology of the harvested cells was markedly different with respect to conventionally (2D) cultured MSCs. In conclusion, the developed bioink can be used to bioprint structures aimed to improve preconditioning MSCs for therapeutic purposes.

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A versatile microfluidic tool for the 3D culture of HepaRG cells seeded at various stages of differentiation

Scientific Reports ◽

10.1038/s41598-021-92011-7 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Manon Boul ◽

Nassima Benzoubir ◽

Antonietta Messina ◽

Rasta Ghasemi ◽

Ismail Ben Mosbah ◽

...

Keyword(s):

Drug Screening ◽

3D Culture ◽

Adhesion Properties ◽

Heparg Cell ◽

Device Architecture ◽

Heparg Cell Line ◽

Toxicological Studies ◽

Heparg Cells ◽

Different Sources ◽

Culture Of Hepatocytes

AbstractThe development of livers-on-a-chip aims to provide pharmaceutical companies with reliable systems to perform drug screening and toxicological studies. To that end, microfluidic systems are engineered to mimic the functions and architecture of this organ. In this context we have designed a device that reproduces series of liver microarchitectures, each permitting the 3D culture of hepatocytes by confining them to a chamber that is separated from the medium conveying channel by very thin slits. We modified the structure to ensure its compatibility with the culture of hepatocytes from different sources. Our device was adapted to the migratory and adhesion properties of the human HepaRG cell line at various stages of differentiation. Using this device, it was possible to keep the cells alive for more than 14 days, during which they achieved a 3D organisation and acquired or maintained their differentiation into hepatocytes. Albumin secretion as well as functional bile canaliculi were confirmed on the liver-on-a-chip. Finally, an acetaminophen toxicological assay was performed. With its multiple micro-chambers for hepatocyte culture, this microfluidic device architecture offers a promising opportunity to provide new tools for drug screening applications.

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3D culture of murine adipose-derived multipotent mesenchymal stromal cells in hydrogel based on carbomer 974P

Cell and Organ Transplantology ◽

10.22494/cot.v6i2.91 ◽

2018 ◽

Vol 6 (2) ◽

Cited By ~ 1

Author(s):

V. Kyryk ◽

O. Kuchuk ◽

A. Mamchur ◽

A. Ustymenko ◽

T. Lutsenko ◽

...

Keyword(s):

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Culture Media ◽

Three Dimensional ◽

3D Culture ◽

Multipotent Mesenchymal Stromal Cells ◽

Term Survival ◽

Hydrogel Scaffolds ◽

3D Cultivation

Actual issues during tissue regeneration are to ensure the survival of transplanted cells at the site of their application and further activity, especially in case of local pathological alterations such as inflammation and ischemia. For this purpose, the matrices that can not only fill the defects of tissues, but also be scaffolds for cells are developed.The aim of this study was to evaluate the effectiveness of 3D cultivation of murine adipose-derived multipotent mesenchymal stromal cells (MSMCs) in hydrogel based on carbomer 974P.Materials and methods. MSMCs were obtained from the adipose tissue of FVB-Cg-Tg(GFPU)5Nagy/J mice transgenic for GFP gene. The cells were phenotyped by flow cytometry and directly differentiated into osteogenic and adipogenic direction to confirm multipotent phenotype. MMSCs were cultured and directly differentiated into osteogenic direction in three-dimensional hydrogel scaffolds. For hydrogel preparation we used carbomer 974P with composition of glycerol, propylene glycol, triethylamine and agarose in original proportion.Results. The three-dimensional hydrogel based on carbomer 974P for the further engraftment with MMSCs was obtained. Modified protocols for the preparation of hydrogels based on carbomer and agarose and their rehydration by culture media for the 3D cultivation of adipose-derived MMSCs have been developed. The optimal concentration of MSMCs and the injection method for engraftment of hydrogels of the required form and size are selected. It was shown that adipose-derived MMSCs in 3D carbomer hydrogel preserve the potential of directed osteogenic differentiation.Conclusion. Three-dimensional hydrogel based on carbomer 974P is capable to support cells, provide the necessary cytoarchitectonics, maintain intercellular interactions, which can promote further long-term survival and specialization of graft.

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Self-Assembling RADA16-I Peptide Hydrogel Scaffold Loaded with Tamoxifen for Breast Reconstruction

BioMed Research International ◽

10.1155/2017/3656193 ◽

2017 ◽

Vol 2017 ◽

pp. 1-10 ◽

Cited By ~ 5

Author(s):

Huimin Wu ◽

Ting Zhou ◽

Lin Tian ◽

Zhengchao Xia ◽

Feng Xu

Keyword(s):

3D Culture ◽

Fat Absorption ◽

Breast Cancer Patients ◽

Hydrogel Scaffold ◽

Hydrogel Scaffolds ◽

In Vivo Experiments ◽

Self Assembling ◽

Apoptosis Related Protein ◽

Mcf 7

More and more breast cancer patients prefer autologous fat tissue transfer following lumpectomy to maintain perfect female characteristics. However, the outcome was not satisfactory due to the transplanted fat absorption. In this study, we prepared two RADA16-I peptide scaffolds with and without tamoxifen. Both scaffolds were transparent, porous, and hemisphere-shaped. The hADSCs isolated from liposuction were attached to the scaffold. The growth inhibition of the hADSCs induced by TAM in 2-demensional (2D) culture was higher than that in TAM-loaded hydrogel scaffold 3D culture (P<0.05); however, the same outcomes were not observed in MCF-7 cells. Correspondingly, the apoptosis of the hADSCs induced by TAM was significantly increased in 2D culture compared to that in scaffold 3D culture (P<0.05). Yet the outcomes of the aoptosis in MCF-7 were contrary. Apoptosis-related protein Bcl-2 was involved in the process. In vivo experiments showed that both scaffolds formed a round mass after subcutaneous implantation and it retained its shape after being pressed slightly. The implantation had no effect on the weight and activity of the animals. The results suggested that TAM-loaded RADA16-I hydrogel scaffolds both provide support for hADSCs cells attachment/proliferation and retain cytotoxic effect on MCF-7 cells, which might be a promising therapeutic breast tissue following lumpectomy.

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