Application: 3D Cell Culture

3D culture methods

Classical cell culture had a major limitation where cells are only able to grow in 2D, the XY plane, rather than there natural state in 3D. This is where 3D cell culture comes in. In living organisms, cells exist in a 3D form in 3D microenvironments with highly organized cell-cell and cell-matrix interactions and complex transport dynamics for nutrients and cells. This means that standard classical 2D cell culture can be unreliable for modeling in vivo drug efficacy and toxicity, which are crucial for drug discovery processes.

3D spheroids are better representatives of in vivo tissue in terms of cellular communication and the development of extracellular matrices (ECMs). ECMS allow cells to move within their spheroid similar to the way cells would move in living tissue. This means that spheroids are better models for cell migration, differentiation, survival, and growth.

Organoids on the other hand are a miniaturized and simplified version of an organ produced in vitro in three dimensions that shows realistic micro-anatomy. They are derived from one or a few cells from a tissue, embryonic stem cells or induced pluripotent stem cells, which can self-organize in three-dimensional culture owing to their self-renewal and differentiation capacities. The last 10 years have shown great advancements in organoid research with cerebral organoids, gut, liver, thymus lung, kidney and even retinal organoids available now.

Scaffold-free technique

3D cell culture is performed in a bioreactor. Being an ideal partner in cell research, we provide the CERO 3D Incubator & Bioreactor. This revolutionary instrument creating optimal cell culture environment. It offers a special 3D cell culture technology that monitors and controls temperature, pH and carbon dioxide levels. Indeed, this is ideal for stem cells, spheroids, organoids and even tissues.

The CERO 3D Incubator and Bioreactor allows for significant improvement of viability and maturation with minimized apoptosis and necrosis, no shear forces, no requirement for embedding substrate. This indeed means that there is a high level of homogeneity and allowing long-term cultivation for more than one year. Being easy to set-up with simple workflows and minimum hands-on time that can be down to less than 2 minutes per day, the CERO 3D Incubator and Bioreactor significantly reduces your running costs.

Aggregate creation with ultra-low attachment plates

In addition, OLS also provides the CEROplate, with ultra-low attachment that simplifies the process for growing 3D aggregates. It features clear wells with a U-bottom to make sample monitoring simple. The unique well geometry of the microplate aids in the formation of an unattached, uniform, round-shaped, single spheroid or organoid in the center of each well. This allows for long-term 3D cell culture over months. CEROplates are compatible with existing imagers, liquid handling and automated stations allowing for perfect data consistency high-throughput applications.

Publications & Links

“Bradley Justice, Nadia A.Badr, Robin A.Felder (2009)
3D cell culture opens new dimensions in cell-based assays
Drug Discovery Today 102-107 ”
DOI: 10.1016/j.drudis.2008.11.006

“Susan Breslin, Lorraine O’Driscoll
Three-dimensional cell culture: the missing link in drug discovery
Drug Discovery Today Volume 240-249”
DOI: 10.1016/j.drudis.2012.10.003

“Simone Allazetta, Tanja C. Hausherr, and Matthias P. Lutolf
Microfluidic synthesis of cell-type-specific artificial extracellular matrix hydrogels
Biomacromolecules 2013, 14, 4, 1122–1131”
DOI: 10.1021/bm4000162″

“Maria Mrakovcic, Markus Absenger, Regina Riedl, Claudia Smole, Eva Roblegg, Leopold F Fröhlich, Eleonore Fröhlich
Assessment of long-term effects of nanoparticles in a microcarrier cell culture system
PLoS One. 2013; 8(2);e56791. ”
DOI: 10.1371/journal.pone.0056791

“Zhen-Yu Yang, Jun-Tao Kan, Ze-Yu Cheng, Xian-Li Wang, Yi-Zhun Zhu & Wei Guo
Daphnoretin-induced apoptosis in HeLa cells: a possible mitochondria-dependent pathway
Cytotechnology volume 66, pages51–61(2014)”
DOI: 10.1007/s10616-013-9536-8

“Bernhard Ellinger, Johanna Silber, Anjali Prashar, Johannes Landskron, Jonas Weber, Sarah Rehermann, Franz-Josef Müller, Stephen Smith, Stephen Wrigley, Kjetil Taskén, Philip Gribbon, Antje Labes, and Johannes F. Imhoff
A phenotypic screening approach to identify anticancer compounds derived from marine fungi
Assay Drug Dev Technol. 2014 Apr 1; 12(3): 162–175 ”
DOI: 10.1089/adt.2013.564

“Ching-Yu Lin, Chi-Hui Huang, Yuan-Kun Wu, Nai-Chen Cheng & Jiashing Yu
Maintenance of human adipose derived stem cell (hASC) differentiation capabilities using a 3D culture
Biotechnology Letters volume 36, pages1529–1537(2014) ”
DOI: 10.1007/s10529-014-1500-y

“Simon Wuest, Stéphane Richard, Isabelle Walther, Reinhard Furrer, Roland Anderegg, Jörg Sekler & Marcel Egli
A Novel Microgravity Simulator Applicable for Three-Dimensional Cell Culturing
Microgravity Science and Technology volume 26, pages77–88(2014)”
DOI: 10.1007/s12217-014-9364-2

“Annette Geltmeier, Beate Rinner, Dennis Bade, Katharina Meditz,Reiner Witt, Uwe Bicker, Catrin Bludszuweit-Philipp, Patrick Maier
Characterization of Dynamic Behaviour of MCF7 and MCF10A Cells in Ultrasonic Field Using Modal and Harmonic Analyses
PLOS ONE”
DOI:10.1371/journal.pone.0134999

“Andreas Elanzew, Annika Sommer, Annette Pusch-Klein, Oliver Brüstle and Simone Haupt
A reproducible and versatile system for the dynamic expansion of human pluripotent stem cells in suspension
Biotechnol. J. 2015, 10, 1589–1599”
DOI: 10.1002/biot.201400757

“Akvilė Jarmalavičiūtė, Virginijus Tunaitis, Ugnė Pivoraitė, Algirdas Venalis, Augustas Pivoriūnas
Exosomes from dental pulp stem cells rescue human dopaminergic neurons from 6-hydroxy-dopamine–induced apoptosis
Cytotherapy Volume 17, Issue 7, July 2015, Pages 932-939”
DOI: 10.1016/j.jcyt.2014.07.013

“Aletta Schnitzler, Anjali Verma, Daniel E. Kehoe, Donghui Jing, Julie R. Murrell, Kara A. Der, Manjula Aysola, Peter J. Rapiejko, Sandhya Punreddy, Martha S. Rook
Bioprocessing of human mesenchymal stem/stromal cells for therapeutic use: Current technologies and challenges
Biochemical Engineering Journal Volume 108, 15 April 2016, Pages 3-13”
DOI: 10.1016/j.bej.2015.08.014

“Guoliang Meng, Shiying Liu, Anna Poon, and Derrick E. Rancourt
Optimizing Human Induced Pluripotent Stem Cell Expansion in Stirred-Suspension Culture
Stem Cells and Development Vol. 26, No. 24”
DOI: 10.1089/scd.2017.0090

“Yongxin Zhang, Xianghan Wang, Mao Pong, Liang Chen, Zhijia Ye
Application of Bioreactor in Stem Cell Culture
J. Biomedical Science and Engineering, 2017, Vol. 10, (No. 11), pp; 485-499”
DOI: 10.4236/jbise.2017.1011037

“Michael Gepp, Benjamin Fischer, André Schulz, Johanne Dobringer, Luca Gentile, Julio A. Vásquez, Julia C. Neubauer, Heiko Zimmermann
Bioactive surfaces from seaweed-derived alginates for the cultivation of human stem cells
Journal of Applied Phycology volume 29, pages2451–2461(2017)”
DOI https://doi.org/10.1007/s10811-017-1130-6

“Alina Čebatariūnienė, Akvilė Jarmalavičiūtė, Virginijus Tunaitis, Alina Pūrienė, Algirdas Venalis, Augustas Pivoriūnasa
Microcarrier culture enhances osteogenic potential of human periodontal ligament stromal cells
Journal of Cranio-Maxillofacial Surgery Volume 45, Issue 6, June 2017, Pages 845-854”
DOI: 10.1016/j.jcms.2017.03.009

“Benjamin Fischer, Anna Meier, Annika Dehne, Aseem Salhotra, Thao Anh Tran, Sascha Neumann, Katharina Schmidt, Ina Meiser, Julia C. Neubauer, Heiko Zimmermann, Luca Gentile
A complete workflow for the differentiation and the dissociation of hiPSCderived cardiospheres
Stem Cell Research 32 (2018) 65–72”
DOI: 10.1016/j.scr.2018.08.015

“Róbert Alföldi, József Á. Balog, Nóra Faragó, Miklós Halmai, Edit Kotogány, Patrícia Neuperger, Lajos I. Nagy, Liliána Z. Fehér, Gábor J. Szebeni andLászló G. Puskás
Single Cell Mass Cytometry of Non-Small Cell Lung Cancer Cells Reveals Complexity of In Vivo and Three-Dimensional Models over the Petri-Dish Cells 2019, 8(9), 1093”
DOI: 10.3390/cells8091093

“Alina Čebatariūnienė, Karolina Kriaučiūnaitė, Justina Prunskaitė, Virginijus Tunaitis, and Augustas Pivoriūnas
Extracellular Vesicles Suppress Basal and Lipopolysaccharide-Induced NFκB Activity in Human Periodontal Ligament Stem Cells
Stem Cells Dev. 2019 Aug 1;28(15):1037-1049.”

“Shamsul Sulaiman, Shiplu Roy Chowdhury, Mh Busra Fauzi, Rizal Abdul Rani,Nor Hamdan Mohamad Yahaya, Yasuhiko Tabata, Yosuke Hiraoka, Ruszymah Binti Haji Idrus and Ng Min Hwei
3D Culture of MSCs on a Gelatin Microsphere in a Dynamic Culture System Enhances Chondrogenesis
Int. J. Mol. Sci. 2020, 21(8), 2688”
DOI: 10.3390/ijms21082688

“Emily Fang, Tana Siboonruang, Jonathan Pache, Eugene Reznikov, Reilly Cashmore
Cell Therapy for Spinal Cord Injuries: Design of a Commercial Manufacturing Facility”
https://sites.duke.edu/emilyfang/files/2020/04/FINAL-REPORT.pdf

“Linda Elberskirch , Thorsten Knoll , Rebecca Konigsmark ¨ , Janis Renner, Nadine Wilhelm ,
Hagen von Briesen , Sylvia Wagner
Microfluidic 3D intestine tumor spheroid model for efficient in vitro investigation of nanoparticular formulations
Journal of Drug Delivery Science and Technology 63 (2021) 102496”
DOI: 10.1016/j.jddst.2021.102496

“Eleonore FRÖHLICH, Claudia MEINDL, Maria MRAKOVCIC, Gerd LEITINGER,
Ramona BAUMGARTNER, Eva ROBLEGG
IN-VITRO TESTING FOR IDENTIFICATION OF LONG-TERM EFFECTS OF NANOPARTICLES
Nanocon 2014 in Brno, CZ”

“S Allazetta, L Kolb, S Zerbib, J Bardy, MP Lutolf
Cell-Instructive Microgels with Tailor-Made Physicochemical Properties
small 2015, 11, No. 42, 5647–5656”
DOI: 10.1002/smll.201501001

“Linda Elberskirch, Ronan Le Harzic, Dietrich Scheglmann, Gerhard Wieland, Arno Wiehe, Maria Mathieu-Gaedke, Hartwig R.A. Golf, Hagen von Briesen, Sylvia Wagner
A HET-CAM based vascularized intestine tumor model as a screening platform for nano-formulated photosensitizers
European Journal of Pharmaceutical Sciences 168 (2022) 106046 ”
DOI: 10.1016/j.ejps.2021.106046

“Pascal Baumann, Jürgen Hubbuch
Downstream process development strategies for effective bioprocesses: Trends, progress, and combinatorial approaches
Eng Life Sci. 2017 Nov; 17(11): 1142–1158.”
DOI: 10.1002/elsc.201600033

“Thao Thi Thanh Ngo, Bella Rossbach, Isabelle Sébastien, Julia C. Neubauer, Andreas Kurtz, Krithika Hariharan
Functional differentiation and scalable production of renal proximal tubular epithelial cells from human pluripotent stem cells in a dynamic culture system Cell Prolif. 2022 Mar; 55(3): e13190”
DOI: 10.1111/cpr.13190

“Stange, Katja, Amir Keric, Andreas Friese, and Monika Röntgen.
Preparation of Spheroids from Primary Pig Cells in a Mid-Scale Bioreactor Retaining Their Myogenic Potential
Cells 2022; 11, no. 9: 1453.”
DOI: 10.3390/cells11091453