Effect of Vascular Formed Endothelial Cell Network on the Invasive Capacity of Melanoma Using the In Vitro 3D Co-Culture Patterning Model

Yamamoto, Shuhei; Hotta, Michael Masakuni; Okochi, Mina; Honda, Hiroyuki

doi:10.1371/journal.pone.0103502

Cited by 23 publications

(15 citation statements)

References 41 publications

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“…We also confirmed that the viability of human cancer cells labeled with MCLs is not decreased by the capture and filtering process (98.8% ± 1.1% before and 97.6% ± 0.9% after) . Furthermore, several research groups, including our group, showed that MCLs do not affect the viability, proliferation, or differentiation of other cell types within the range of magnetite concentration tested . According to these observations, the viability and proliferation of CTCs in clinical samples should not be inhibited by MCLs under the conditions used in the present study.…”

Section: Discussionsupporting

confidence: 87%

“…A pin‐holder device was used to position the captured CTCs on the culture dish with or without a fibroblast feeder layer, as described previously . A culture dish with or without the feeder layer was placed on the pin‐holder device with a magnet; to this dish, we added the whole recovered DMEM containing CTCs in order to seed the magnetically labeled CTCs.…”

Section: Methodsmentioning

confidence: 99%

“…Here, we report a novel method for ex vivo culture of CTCs using a fibroblast feeder layer and a magnetic‐force‐based cell coculture protocol . A fibroblast feeder layer has long been used for cultivation of stem cells, including embryonic stem cells and induced pluripotent stem cells .…”

Section: Introductionmentioning

confidence: 99%

“…Therefore, we hypothesized that coculture of isolated CTCs with fibroblasts would promote proliferation of the CTCs. Furthermore, to facilitate attachment of the isolated CTCs to the fibroblast feeder layer, we used the magnetic‐force‐based coculture method . In this method, magnetically labeled cells are positioned on the cultured feeder layer by a magnetic field.…”

Section: Introductionmentioning

confidence: 99%

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Ex vivo culture of circulating tumor cells using magnetic force‐based coculture on a fibroblast feeder layer

Yamamoto

Shimizu

Fei

et al. 2016

Biotechnology Journal

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Phenotype-based analysis of circulating tumor cells (CTCs) is a promising approach to identification of new therapeutic targets and to elucidation of the biological properties. Nonetheless, ex vivo culturing of CTCs is still a technical challenge. Here, we develop a novel ex vivo culture method for CTCs using a fibroblast feeder layer and a magnetic coculture protocol. CTCs in the blood of a mouse metastasis model are labeled magnetically with magnetite nanoparticles. The labeled CTCs are isolated by a magnetic capture column and a size-selective capture filter. The isolated CTCs are positioned on a fibroblast feeder layer by the magnetic force. As a result, we observe adhesion and proliferation of the CTCs under the conditions of the fibroblast feeder layer and the magnetic force, whereas no adhesion or proliferation is observed without the feeder layer. After that, we culture the CTCs and obtain three CTC-derived cell lines. Using these cell lines, we perform phenotype-based analyses of invasiveness and drug resistance and find that the CTC-derived cell lines are more malignant than the original cells. Thus, the proposed method would be a promising approach to ex vivo culture of CTCs for phenotype-based analysis, and possibly used in cancer treatment.

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Section: Discussionsupporting

confidence: 87%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Ex vivo culture of circulating tumor cells using magnetic force‐based coculture on a fibroblast feeder layer

Yamamoto

Shimizu

Fei

et al. 2016

Biotechnology Journal

Self Cite

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“…We have developed a cell patterning method for mammalian cells using magnetite nanoparticles and the pin-holder magnetic device for single cell analysis (Ino et al, 2008;Okochi et al, 2013a;Okochi et al, 2013b;Okochi et al, 2009;Yamamoto et al, 2014). In this method, we used magnetite cationic liposomes (MCLs), which are cationic liposomes containing 10 nm-magnetite nanoparticles.…”

Section: Introductionmentioning

confidence: 99%

A single cell culture system using lectin‐conjugated magnetite nanoparticles and magnetic force to screen mutant cyanobacteria

Arai

Okochi

Shimizu

et al. 2015

Biotech & Bioengineering

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Cyanobacteria can be utilized as a potential biocatalyst for the production of biofuels and biochemicals directly from CO2. Useful mutants of cyanobacteria, which can grow rapidly or are resistant to specific metabolic products, are essential to improve the productivity of biofuels. In this study, we developed a single cell culture system to effectively screen mutant cyanobacteria using magnetite nanoparticles and magnetic force. Lens culinaris Agglutinin (LCA) was selected as a lectin, which binds to the surface of Synechococcus elongatus PCC7942 cells and the LCA-conjugated magnetite cationic liposomes (MCLs) were developed for magnetic labeling of PCC7942 cells. The MCL-labeled PCC7942 cells were magnetically patterned at a single cell level by using 6,400 iron pillars of the pin-holder device. The device enabled 1,600 single cells to be arrayed in one square centimeter. We cultured the patterned cells in liquid medium and achieved higher colony-forming ratio (78.4%) than that obtained using conventional solid culture method (4.8%). Single cells with different properties could be distinguished in the single cell culture system depending on their growth. Furthermore, we could selectively pick up the target cells and subsequently perform efficient isolation culture. The ratio of successful isolation culture using the developed method was 13 times higher than that of the conventional methods. Thus, the developed system would serve as a powerful tool for screening mutant cyanobacteria.

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High‐Precision 3D Bio‐Dot Printing to Improve Paracrine Interaction between Multiple Types of Cell Spheroids

Jeon

Heo

Kim

et al. 2020

Adv Funct Materials

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Cell-cell interaction accounts for one of the most influential factors affecting the viability and functionality of cell-based tissue models. In this respect, various methods capable of producing micro-patterns with cell spheroids are introduced to simultaneously improve contact-dependent and-independent cell-cell interactions. However, no method has yet been designed to effectively generate precise 3D patterns with multiple spheroid types. In this study, a new high-precision and convenient 3D spheroid printing technology is developed, designated as 3D bio-dot printing. This new technique is designed to produce cell-laden, non-adhesive micro-pores within 3D structures to allow cell spheroids to be induced at printed sites. Experimental results show that various cell types, including hepatocytes, pancreatic β-cells, and breast cancer cells, can be employed for the in situ formation of cell spheroids, and 3D freeform structures with multiple spheroid types can be printed. Moreover, this novel technology can also be used for performing 3D invasion assays. More importantly, it ensures that the precise control of spheroid size and position is achieved at micrometer scale. Finally, the usefulness of this novel technology is demonstrated by producing multicellular micro-patterns with primary hepatocyte spheroids and endothelial cells, that exhibit significantly improved long-term hepatic function and drug metabolism.

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Effect of Vascular Formed Endothelial Cell Network on the Invasive Capacity of Melanoma Using the In Vitro 3D Co-Culture Patterning Model

Cited by 23 publications

References 41 publications

Ex vivo culture of circulating tumor cells using magnetic force‐based coculture on a fibroblast feeder layer

Ex vivo culture of circulating tumor cells using magnetic force‐based coculture on a fibroblast feeder layer

A single cell culture system using lectin‐conjugated magnetite nanoparticles and magnetic force to screen mutant cyanobacteria

High‐Precision 3D Bio‐Dot Printing to Improve Paracrine Interaction between Multiple Types of Cell Spheroids

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