​アプリケーション

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​ラベルフリーと3次元培養

3次元培養細胞

(シングルスフェロイド)

Cell3iMager duosで撮像

3次元培養細胞

(マルチスフェロイド)

Cell3iMager duosで撮像

3次元培養細胞

(スフェロイド)

Cell3iMager Estierで撮像

ラベルフリーで広がるアッセイ

 培養細胞を利用した研究では、蛍光解析を行うことにより、様々な情報を取得することが出来ます。その一方、蛍光解析は、研究に様々な制限を与える場合があります。例えば、蛍光染色は、固定化から観察まで、手間とコストが掛かるばかりでなく、培養経過に伴う細胞の動的変化を観察するのは困難です。また、強制発現した蛍光タンパク質は細胞の基底状態に影響を与える可能性があります。昨今、大きく発展している再生医療研究やがん研究では、細胞のダイナミズムを研究するケースも増大しています。これら研究の為、基底状態の細胞を生きたまま観察、管理する必要性は益々高まっています。ラベルフリーアッセイを導入することで、生きた細胞をそのまま解析出来るばかりでなく、高価な蛍光試薬や遺伝子導入試薬を用いる必要がありません。Cell3iMagerシリーズによるラベルフリーアッセイは、これら研究への利用に最適なシステムです。

3次元培養の可能性

 生物の組織は、複雑な立体構造を持ちます。vitroの系を用いて生命現象を解析する為には、平面培養に比べ、生体により近い構造を模した3次元培養での検討が重要となります。例えば、再生医療研究やがん研究では、生体により近い特徴を持つ3次元培養細胞を用いて研究を行うことが、今後益々必要になっていくと予想されています。

Cell3iMagerシリーズによる研究への貢献

 Cell3iMagerシリーズは、3次元培養のオルガノイドやスフェロイドについて、3次元撮像、定量解析を行う、ラベルフリーセルイメージャーです。特に、Cell3iMagerのシステム化されたワークフローは、研究のスループットと再現性を向上させます。

​再生医療と創薬を支える基盤技術「三次元培養細胞」

​アプリケーションノート

共同研究

       高速細胞スキャナーを用いたスフェロイド培養

       による抗がん剤スクリーニング法の開発 (CC-5000)

        (長浜バイオ大学 水上研究室との共同研究)

 

       迅速な胚様体(EB)の作製とサイズ評価 (CC-5000)

        (InSphero AG社との共同研究)

       複雑な3D MCTSモデルの研究に最適な高速3D

       細胞スキャナー (CC-5000)

        (MD Anderson Cancer Centerとの共同研究)

Cell3iMager シリーズ使用文献

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) 

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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