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Applications

Column

​Label-free and 3D cell culture

Label-free assay

　Laboratory research and applications have always benefitted by the utility of fluorescence based probes to evaluate and markers or processes for better understanding of processes. As beneficial these have been in life sciences, these also pose limitations on assessing changes at cellular/molecular levels , because, fluorescence staining is not only labor-intensive and costly, from fixation to observation, but also makes it difficult to observe the dynamic changes occurring in cells over the course of culture. In addition, exogenous expression of fluorescent protein may affect the ground state of cells. In recent years, immense advances have been made in the area of regenerative medicine and cancer research. These advances have been possible by  assessing the changes at cellular or molecular level by studying dynamic changes in cells. To this end, there is an ever-increasing need to observe and manage cellular changes without having the need to exogenously introduce markers tagged with fluorescent probes of interest or introduce fluorescent probes into the cells. Label-free assays have, therefore, rapidly grown to be of high interest in research and applications. SCREEN imaging technology, Cell3iMager series have proven to be  ideal systems in research and development for performing label-free assays.

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Possibility of 3D cell culture

　There has been rapid advance in in vitro and ex vivo based laboratory research with 3D cell culture gaining relevance in cell based assays. It has already been shown that 3D cell cultures reflect in vivo environment, therefore, highly important in assessing the phenotypic changes. It is also well known that 3D cell cultures pose a complicated three-dimensional structure, in order to analyze biological phenomena using the in vitro system, it is important to utilize technology to evaluate the complex 3D structures as well as tissues. Over the past years, 3D cell culture platforms are being used in disease modelling drug discovery, early development. Key areas such as , regenerative medicine and cancer research are utilizing three-dimensional cell culture assays to better understand the biological processes.

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Contributions to Research with our Cell3iMager Series

　The Cell3iMager series specializes in label-free 3D imaging and quantitative analysis of 3D cultured organoids and spheroids. In particular, the versatile and robust Cell3imajer technology  exhibits key aspects in useful in high throughput drug discovery and early development.

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​3D-cultured cells, a fundamental technology that supports regenerative medicine and drug discovery research

3D Culture Cells

(Single spheroid)
Imaging with Cell3iMager duos

3D Culture Cells

(Multi spheroid)
Imaging with Cell3iMager duos

3D Culture Cells

(Single spheroid)
Imaging with Cell3iMager Estier

​Application note

  1. 3D cell culture by hanging drop ​

  2. Colony-forming units assay

  3. 3D cell culture in GravityTrap

  4. 3D Cell Culture by using hydrogels

  5. Pancreatic β cells stained by DTZ

Collaboration

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"Development of spheroid-based anticancer drug screening method using high-speed cell culture scanner" (CC-5000)

(Collaborative research with Nagahama Institute of Bio-Science and Technology Mizukami Laboratory)

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"Rapid production and size assessment of Embryoid Bodies ( EBs )" (CC-5000)

(Collaborative research with InShero AG)

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"High-Speed 3D Cell Scanner —Perfect for the Study of Complex 3D MCTS Models" (CC-5000)

(Collaborative research with MD Anderson Cancer Center)

  1. Cell Viability and Cytotoxicity Assay

  2. Single cell cloning hybridoma

  3. Single cell cloning

  4. 3D Cell Culture by using FCeM

  5. Assessing the growth of 3D spheroids cultured in Matrigel

  6. Analysis of microscope images

  7. Counting fluorescence stained adherent cells

  8. Evaluation of hypoxia level in 2D/3D-cultured cells

  9. Cell Proliferation

  10. Label free colony-forming units assay

  11. Confluency

  12. Neuronal differentiation

  13. Suspension cultured cell

  14. Tissue sections

  15. Label free analysis of spheroids in microwells

  16. Deep Learning for cell classification

  17. Analysis of spheroids in microwells

  18. Organelle Staining

  19. Aniti-Cancer Drug Screening

  20. LiveDead Assay

  21. Invasion Assay

  22. Label free analysis of spheroids in microwells

  1. Real volume of spheroids

  2. Thickness of Cell Sheet

Publications

1.

Visualizing dynamics of angiogenic sprouting from a three-dimensional microvasculature model using stage-top optical coherence tomography 

Haruko Takahashi, Keisuke Kato, Kenji Ueyama, Masayoshi Kobayashi, Gunwoong Baik,Yasuhiro Yukawa, Jun-ichi Suehiro, Yukiko T. Matsunaga. 

Scientific reports, (2017), 7:42426.

2.

A Vascular Endothelial Growth Factor-Dependent Sprouting Angiogenesis Assay Based on an In Vitro Human Blood Vessel Model for the Study of Anti-Angiogenic Drugs 

Joris Pauty, Ryo Usuba, Irene Gayi Cheng, Louise Hespel, Haruko Takahashi, Keisuke Kato, Masayoshi Kobayashi, Hiroyuki Nakajima, Eujin Lee, Florian Yger, Fabrice Soncin, Yukiko T. Matsunaga. 

EBioMedicine, (2018), 27: 225–236. 

3.

A 3D in vitro pericyte-supported microvessel model: visualisation and quantitative characterisation of multistep angiogenesis 

Eujin Lee, Haruko Takahashi, Joris Pauty, Masayoshi Kobayashi, Keisuke Kato, Maki Kabara, Jun-ichi Kawabe and Yukiko T. Matsunaga. 

Journal of Materials Chemistry B, (2018), 6; 1085-1094. 

4.

EGFL7 regulates sprouting angiogenesis and endothelial integrity in a human blood vessel model 

Ryo Usuba, Joris Pauty, Fabrice Soncin, and Yukiko T.Matsunaga 

Biomaterials, Volume 197, March 2019, Pages 305-316

5.

Directed differentiation of human induced pluripotent stem cells into mature stratified bladder urothelium 

Kotaro Suzuki, Michiyo Koyanagi-Aoi, Keiichiro Uehara, Nobuyuki Hinata, Masato Fujisawa & Takashi Aoi 

Scientific Reports volume 9, Article number: 10506 (2019) 

6.

A simple three-dimensional gut model constructed in a restricted ductal microspace induces intestinal epithelial cell integrity and facilitates absorption assays 

Tadaaki Nakajima, Katsunori Sasaki, Akihiro Yamamori, Kengo Sakurai, Kaori Miyata, Tomoyuki Watanabe and Yukiko T Matsunaga   

Biomater. Sci., 2020, First published:31 Aug 2020 

1.

Fetal bovine serum enlarges the size of human pancreatic cancer spheres accompanied by an increase in the expression of cancer stem cell markers 

Norihiko Sasaki, Masashi Toyoda, Fumio Hasegawa, Masakazu Fujiwara, Fujiya Gomi b, Toshiyuki Ishiwata. 

Biochemical and Biophysical Research Communications, Vol. 514, Issue 1, 18 June 2019, Pages 112-117. 

2.

Ganglioside GM2, highly expressed in the MIA PaCa-2 pancreatic ductal adenocarcinoma cell line, is correlated with growth, invasion, and advanced stage 

Norihiko Sasaki, Kenichi Hirabayashi, Masaki Michishita, Kimimasa Takahashi, Fumio Hasegawa, Fujiya Gomi, Yoko Itakura, Naoya Nakamura, Masashi Toyoda,Toshiyuki Ishiwata 

Scientific Reports volume 9, Article number: 19369 (2019) 

3.

Hydrodynamic stress stimulates growth of cell clusters via the ANXA1/PI3K/AKT axis in colorectal cancer 

Takeshi Hagihara, Jumpei Kondo, Hiroko Endo, Masayuki Ohue, Yoshiharu Sakai and Masahiro Inoue 

Scientific Reports volume 9, Article number: 20027 (2019) 

4.

Novel bone microenvironment model of prostate cancer with chitosan fiber matrix and osteoblast in 3D culture 

Masahiro Samoto, Hiroaki Matsumoto, Hiroshi Hirata, Sho Ozawa, Junichi Mori, Ryo Inoue, Seiji Yano, Yoshiaki Yamamoto and Hideyasu Matsuyama 

AACR 10.1158/1538-7445.AM2020-321 Published August 2020 

5.

The anti-inflammatory effect of the gut lactic acid bacteria-generated metabolite 10-oxo-cis-6,trans-11-octadecadienoic acid on monocytes 

Shuhei Hagiwara, Kazuki Nagata, Kazumi Kasakura, Fumiya Sakata, Shigenobu Kishino, Jun Ogawa, Takuya Yashiro and Chiharu Nishiyama 

BBRC, Volume 530, Issue 1, 10 September 2020, Pages 342-347

6.

Methods for Automated Single Cell Isolation and Sub‐Cloning of Human Pluripotent Stem Cells 

Valeria Fernandez Vallone  Narasimha Swamy Telugu  Iris Fischer  Duncan Miller  Sandra Schommer  Sebastian Diecke  Harald Stachelscheid

Current Protocols in Stem Cell Biology,Volume55, Issue1,December 2020,e123　First published: 21 September 2020

7.

Generation of human induced pluripotent stem cell-derived liver buds with chemically defined and animal origin-free media

Keisuke Sekine, Shimpei Ogawa, Syusaku Tsuzuki, Tatsuya Kobayashi, Kazuki Ikeda, Noriko Nakanishi, Kenta Takeuchi, Eriko Kanai, Yugo Otake, Satoshi Okamoto, Tsuyoshi Kobayashi, Takanori Takebe & Hideki Taniguchi 

Scientific Reports volume 10, Article number: 17937 (2020) 

8.

Functional analysis of isoflavones using patient-derived human colonic organoids

MaoTsuchiya,GoIto,MinamiHama,SayakaNagata,AmiKawamoto,KoheiSuzuki,HiromichiShimizu,ShoAnzai,JunichiTakahashi,ReikoKuno,SayakaTakeoka,YuiHiraguri,HadyYukiSugihara,TomohiroMizutani,ShiroYui,ShigeruOshima,KiichiroTsuchiya,MamoruWatanabe,RyuichiOkamoto

Biochemical and Biophysical Research Communications, Volume 542, 26 February 2021, Pages 40-47

9.

Nickel ions attenuate autophagy flux and induce transglutaminase 2 (TG2) mediated post-translational modification of SQSTM1/p62

Emi Aonuma, Akiko Tamura, Hiroki Matsuda, Takehito Asakawa, Yuriko Sakamaki, Kana Otsubo, Yoichi Nibe, Michio Onizawa, Yasuhiro Nemoto, Takashi Nagaishi, Kiichiro Tsuchiya, Tetsuya Nakamura, Motohiro Uo, Mamoru Watanabe, Ryuichi Okamoto, Shigeru Oshima

Biochem Biophys Res Commun. 2021 Jan 19;542:17-23. doi: 10.1016/j.bbrc.2021.01.023.

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1.

Phenotypic Drug Discovery in 3D, Testing Drug Efficacy by Rapid Size ProfilingOver Time with Tumor Microtissues 

Randy Strube, Ph.D., Andreia F.Fernandes, Markus Furter, Jens M. Kelm,Ph.D., and David A. Fluri, Ph.D. 

Genetic Engineering & Biotechnology News, (2014), Vol. 34, No. 16. 

2.

Human Dynactin-Associated Protein Transforms NIH3T3 Cells to Generate Highly Vascularized Tumors with Weak Cell-Cell Interaction 

Tatsuki Kunoh, Weixiang Wang, Hiroaki Kobayashi, Daisuke Matsuzaki, Yuki Togo, Masahiro Tokuyama, Miho Hosoi, Koichi Koseki, Shu-ichi Wada, Nobuo Nagai, Toshinobu Nakamura, Shintaro Nomura, Makoto Hasegawa, Ryuzo Sasaki, Tamio Mizukami. 

PLOS ONE, (2015), August 18. 

3.

Three-Dimensional Spheroid Cell Culture Model for Target Identification Utilizing High-Throughput RNAi Screens 

LaKesla R. Iles, Geoffrey A. Bartholomeusz. 

High-Throughput RNAi Screening (2016), 21-135.

4.

PGE2 is a direct and robust mediator of anion/fluid secretion by human intestinal epithelial cells 

Satoru Fujii, Kohei Suzuki, Ami Kawamoto, Fumiaki Ishibashi, Toru Nakata, Tatsuro Murano, Go Ito, Hiromichi Shimizu, Tomohiro Mizutani, Shigeru Oshima, Kiichiro Tsuchiya, Tetsuya Nakamura, Akihiro Araki, Kazuo Ohtsuka, Ryuichi Okamoto,  Mamoru Watanabe. 

Scientific Reports, (2016), 36795. 

5.

PAF promotes stemness and radioresistance of glioma stem cells 

Derrick Sek Tong Ong, Baoli Hu, Yan Wing Ho, Charles-Etienne Gabriel Sauvé, Christopher A. Bristow, Qianghu Wang, Asha S. Multani, Peiwen Chen, Luigi Nezi, Shan Jiang, Claire Elizabeth Gorman, Marta Moreno Monasterio, Dimpy Koul, Matteo Marchesini, Simona Colla, Eun-Jung Jin, Erik P. Sulman, Denise J. Spring, Wai-Kwan Alfred Yung, Roel G. W. Verhaak, Lynda Chin, Y. Alan Wang, and Ronald A. DePinho 

PNAS October 24, 2017 114 (43) 

6.

Single cell analysis of Crohn’s disease patient-derived small intestinal organoids reveals disease activity-dependent modification of stem cell properties 

Kohei Suzuki, Tatsuro Murano, Hiromichi Shimizu, Go Ito, Toru Nakata, Satoru Fujii, Fumiaki Ishibashi, Ami Kawamoto, Sho Anzai, Reiko Kuno, Konomi Kuwabara, Junichi Takahashi, Minami Hama, Sayaka Nagata, Yui Hiraguri, Kento Takenaka, Shiro Yui, Kiichiro Tsuchiya, Tetsuya Nakamura, Kazuo Ohtsuka, Mamoru Watanabe, Ryuichi Okamoto. 

Journal of Gastroenterology, (2018), 1–13. 

7.

Sertraline, chlorprothixene, and chlorpromazine characteristically interact with the REST-binding site of the corepressor mSin3, showing medulloblastoma cell growth inhibitory activities 

Jun-ichi Kurita, Yuuka Hirao, Hirofumi Nakano, Yoshifumi Fukunishi & Yoshifumi Nishimura. 

Scientific Reports 8, Article number: 13763 (2018). 

8.

Up-regulation of Bone Morphogenetic Protein 7 by 2-Hydroxycinnamaldehyde Attenuates HNSCC Cell Invasion 

SUNG-MIN KANG1, JINKYUNG KIM1, SOO HYUN KANG1, SU YOUNG OH1, HEON-JIN LEE1, BYOUNG-MOG KWON2 and SU-HYUNG HONG1 

Anticancer Research October 2018 vol. 38 no. 10 5747-5757 

9.

Acquired JHDM1D–BRAF Fusion Confers Resistance to FGFR Inhibition in FGFR2-Amplified Gastric Cancer 

Hitoshi Sase, Yoshito Nakanishi, Satoshi Aida, Kana Horiguchi-Takei, Nukinori Akiyama, Toshihiko Fujii, Kiyoaki Sakata, Toshiyuki Mio, Masahiro Aoki and Nobuya Ishii 

Molecular cancer theraputics-17-1022 Published October 2018 

10.

Automated, high-throughput measurement of size and growth curves of small organisms in well plates 

James Duckworth, Tjalling Jager & Roman Ashauer 

Scientific Reportsvolume 9, Article number: 10 (2019)  

11.

A novel electron emission-based cell culture device promotes cell proliferation and differentiation of pre-osteoblastic MC3T3-E1 cells 

Fumiaki Sugimori, Hiroyuki Hirakawa, Ai Tsutsui, Hiroyuki Yamaji, Shohei Komaru, Mai Takasaki, Tadashi Iwamatsu, Toshimasa Uemura, Yo Uemura, Kenichi Morita, Takashi Tsumura 

PLOS, March 28, 2019 

12.

Monospecific antibody targeting of CDH11 inhibits epithelial-to-mesenchymal transition and represses cancer stem cell-like phenotype by up-regulating miR-335 in metastatic breast cancer, in vitro and in vivo 

Jia-Hong Chen, Wen-Chien Huang, Oluwaseun Adebayo Bamodu, Peter Mu-Hsin Chang, Tsu-Yi Chao & Tse-Hung Huang 

BMC Cancer volume 19, Article number: 634 (2019) 

13.

Wild-type TP53 defined gamma-secretase inhibitor sensitivity and synergistic activity with doxorubicin in GSCs 

Chen Zhang, Emmanuel Martinez-Ledesma, Feng Gao, Wei Zhang, Jie Ding, Shaofang Wu, Xiaolong Li, Jimin Wu, Ying Yuan, Dimpy Koul, and WK Alfred Yung 

Am J Cancer Res. 2019; 9(8): 1734–1745. 2019 Aug 1. 

14.

A 3D Heterotypic Multicellular Tumor Spheroid Assay Platform to Discriminate Drug Effects on Stroma versus Cancer Cells 

Zoe Weydert, Madhu Lal-Nag, Lesley Mathews-Greiner, Christoph Thiel, Henrik Cordes, Lars Küpfer, Patrick Guye, Jens M. Kelm, Marc Ferrer 

SLAS Volume: 25 issue: 3, page(s): 265-276 October 29, 2019 

15.

Promoter-Level Transcriptome Identifies Stemness Associated With Relatively High Proliferation in Pancreatic Cancer Cells 

Ru Chen, Aiko Sugiyama, Naoyuki Kataoka, Masahiro Sugimoto, Shoko Yokoyama, Akihisa Fukuda, Shigeo Takaishi and Hiroshi Seno 

Frontiers in Oncology Gastrointensitial Cancers, 20 March 2020 

16.

Parallelized Wireless Sensing System for Continuous Monitoring of Microtissue Spheroids 

Lei Dong, Paolo S. Ravaynia, Qing-An Huang, Andreas Hierlemann, and Mario M. Modena 

ACS Sens. 2020, 5, 7, 2036–2043, Publication Date:June 10, 2020 

17.

Preclinical Evaluation of the Novel Small-Molecule MSI-N1014 for Treating Drug-Resistant Colon Cancer via the LGR5/β-catenin/miR-142-3p Network and Reducing Cancer-Associated Fibroblast Transformation 

Vijesh Kumar Yadav, Yan-Jiun Huang, Thomashire Anita Georg, Po-Li Wei, Maryam rachmawati Sumitra, Ching-Liang Ho, Tzu-Hao Chang, Alexander T. H. Wu and Hsu-Shan Huang 

Cancers 2020, 12(6), 1590; Published: 16 June 2020 

18.

Co-Microencapsulation of Islets and MSC CellSaics, Mosaic-Like Aggregates of MSCs and Recombinant Peptide Pieces, and Therapeutic Effects of Their Subcutaneous Transplantation on Diabetes 

Yusuke Mochizuki, Ryo Kogawa, Ryuta Takegami, Kentaro Nakamura, Akira Wakabayashi, Tadashi Ito and Yasuhiro Yoshioka 

Biomedicines 2020, 8(9), 318; Published: 31 August 2020 

19.

PREDICTING METABOLISM-RELATED DRUG-DRUG INTERACTIONS USING A MULTI-TISSUE MICROPHYSIOLOGICAL SYSTEM 

Christian Lohasz, Flavio Bonanini, Kasper Renggli, Olivier Frey, and Andreas Hierlemann. 

ETH Zurich NO. 26595, 2020 

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