Survival and structural evaluations of three-dimensional tissues fabricated by the hierarchical cell manipulation technique

Chetprayoon, Paninee; Kadowaki, Koji; Matsusaki, Michiya; Akashi, Mitsuru

doi:10.1016/j.actbio.2012.08.019

Cited by 30 publications

(19 citation statements)

References 33 publications

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“…Regarding long-term stability, we currently reported one of the most important functions as a tissue, the structural stability of the 3D-constructs [59]. In this paper, we demonstrated that 5L-NHDF tissues maintained their original thickness, living-cell density and layer number even after 1 month of culture.…”

Section: Future Directionsmentioning

confidence: 61%

Three-dimensional cell culture technique and pathophysiology

Matsusaki

Case

Akashi

2014

Advanced Drug Delivery Reviews

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Three-dimensional (3D) tissue constructs consisting of human cells have opened a new avenue for tissue engineering, pharmaceutical and pathophysiological applications, and have great potential to estimate the dynamic pharmacological effects of drug candidates, metastasis processes of cancer cells, and toxicity expression of nano-materials, as a 3D-human tissue model instead of in vivo animal experiments. However, most 3D-cellular constructs are a cell spheroid, which is a heterogeneous aggregation, and thus the reconstruction of the delicate and precise 3D-location of multiple types of cells is almost impossible. In recent years, various novel technologies to develop complex 3D-human tissues including blood and lymph capillary networks have demonstrated that physiological human tissue responses can be replicated in the nano/micro-meter ranges. Here, we provide a brief overview on current 3D-tissue fabrication technologies and their biomedical applications. 3D-human tissue models will be a powerful technique for pathophysiological applications.

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Section: Future Directionsmentioning

confidence: 61%

Three-dimensional cell culture technique and pathophysiology

Matsusaki

Case

Akashi

2014

Advanced Drug Delivery Reviews

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“…The contrast between primary cells and immortalized cell lines is also evident in longer culture periods. For instance, in a 3D model immortalized fibroblasts proliferated excessively and displayed heterogeneous and random increases in thickness of 3D layers, while normal human primary dermal fibroblasts demonstrated consistent DNA levels and maintained consistent thickness for layered structures [54]. Primary cells on the other hand have the the disadvantage of donor-to-donor variability [1] and therefore are not well characterized, making it more difficult to compare outcomes between experiments.…”

Section: Temporal Component Of Tissue Engineeringmentioning

confidence: 99%

Strategies for improving the physiological relevance of human engineered tissues

Abbott

Kaplan

2015

Trends in Biotechnology

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This review examines important robust methods for sustained, steady state, in vitro culture. To achieve ‘physiologically relevant’ tissues in vitro additional complexity must be introduced to provide suitable transport, cell signaling, and matrix support for cells in 3D environments to achieve stable readouts of tissue function. Most tissue engineering systems draw conclusions on tissue functions such as responses to toxins, nutrition or drugs based on short term outcomes with in vitro cultures (2–14 days). However, short term cultures limit insight with physiological relevance, as the cells and tissues have not reached a steady state.

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“…We have developed a simple and unique bottom-up approach, "hierarchical cell manipulation technique," which employs nanometer-sized LbL films consisting of FN and gelatin (G) as a nano-ECM. 14,21,[25][26][27][28][29] The FN-G nanofilms were prepared directly on the cell surface, and we discovered that at least 6 nm thick FN-G films acted as a stable adhesive surface for adhesion of the second cell layer. Various 3D-layered constructs consisting of single or multiple types of cells were successfully fabricated, 28,29 and the higher cellular activities induced from the 3D-structures as compared to monolayer structure were observed.…”

Section: Introductionmentioning

confidence: 99%

“…14,21,[25][26][27][28][29] The FN-G nanofilms were prepared directly on the cell surface, and we discovered that at least 6 nm thick FN-G films acted as a stable adhesive surface for adhesion of the second cell layer. Various 3D-layered constructs consisting of single or multiple types of cells were successfully fabricated, 28,29 and the higher cellular activities induced from the 3D-structures as compared to monolayer structure were observed. 25 The multilayered constructs like a blood vessel wall structure indicated almost the same drug response as in vivo natural blood vessels, suggesting the possibility to use as an in vitro blood vessel model to analyze drug response.…”

Section: Introductionmentioning

confidence: 99%

Construction of three-dimensional liver tissue models by cell accumulation technique and maintaining their metabolic functions for long-term culture without medium change

Matsuzawa

Matsusaki

Akashi

2014

J. Biomed. Mater. Res.

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Three-dimensional (3D) hepatocyte cultures have attracted much attention to obtain high biological functions of hepatocyte for pharmaceutical drug assessment. However, maintaining the high functions for over one month is still a key challenge although many approaches have been reported. In this study, we demonstrate for the first time simple and rapid construction of 3D-hepatocyte constructs by our cell accumulation technique and their high biological functions for one month, without any medium change. The human hepatocyte carcinoma (HepG2) cells were coated with ∼ 7 nm-sized extracellular matrix (ECM) films consisting of fibronectin (FN) and gelatin (G), and then incubated in cell culture insert to construct 3D-tissue constructs for 24 h. The thickness of obtained 3D-HepG2 constructs was easily controlled by altering seeding cell number and the maximum is over 100 μm. When a large volume of culture media was employed, the 3D-constructs showed higher mRNA expression of albumin and some cytochrome P450 (CYP) enzymes as compared to general two-dimensional (2D) culture. Surprisingly, their high cell viabilities (over 80%) and high mRNA expressions were successfully maintained without medium change for at least 27 days. These results demonstrate novel easy and rapid technique to construct 3D-human liver tissue models which can maintain their high functions and viability for 1 month without medium change.

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Survival and structural evaluations of three-dimensional tissues fabricated by the hierarchical cell manipulation technique

Cited by 30 publications

References 33 publications

Three-dimensional cell culture technique and pathophysiology

Three-dimensional cell culture technique and pathophysiology

Strategies for improving the physiological relevance of human engineered tissues

Construction of three-dimensional liver tissue models by cell accumulation technique and maintaining their metabolic functions for long-term culture without medium change

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