Dear All,

Below please find the fifteen issue of the newsletter of the 3D Cell Culture Core (3D3C) Facility of the Birck Nanotechnology Center.  The newsletter is also available online (https://nanohub.org/groups/3d3cfacility/news). 

The five sections in the newsletter are:
3D at Purdue – this section highlights 3D cell culture-based research activity at Purdue
3D in action - this section highlights recent activity in 3D3C
3D in focus – this section presents the current work on a specific 3D cell culture model or technique
3D in publications – this section brings a collection of recent publications on 3D cell culture
3D in meetings – this section includes a list of upcoming meetings related to 3D cell culture

The newsletter will be available every two months.  If you do not wish to receive the 3D3C newsletter in the future, please reply “cancel” to unsubscribe.
Please contact me if you have questions. 


Yours Sincerely,
 
Tim Kwok
Facility Manager
3D Cell Culture Core (3D3C) Facility
Birck Nanotechnology Center
Purdue University

 


 

Volume 15, April 2018


 

 

 

3D at Purdue

3D in Action

3D in Focus

3D in Publications

3D in Meetings




3D at Purdue

 


Gradient-On-a-Chip to Recreate Heterogenous Microenvironments
 

 

The Lelièvre and Ziaie laboratories collaborated to design a gradient-on-a-chip (GOC) system in which cells can be exposed to increasing concentrations of chemicals of interest in the same culture chamber. The GOC is made of multiple parts that can be assembled easily with screws and it is reusable (only the paper insert needs to be changed after each culture) (Figure 1). The microfluidic network includes a mixer to create the gradient (Figure 2).

The idea behind the design of this cell culture device is that tissue heterogeneity controls cell behavior and in diseases like cancers, promotes progression and treatment resistance. Part of the heterogeneity in cell populations originates from the microenvironment in which chemicals exist in various local concentrations and thus, differently influence cells of a same tissue. Another use of the GOC is to establish concentration thresholds at which a chemical or a drug starts having an effect depending on tissue characteristics. For instance, to validate the use of the system, in their collaborative manuscript (Chittiboyina/Rahimi et al., ACS Biomat Sci & Eng, 2018) the Lelièvre and Ziaie teams demonstrated that a higher tissue stiffness (as seen during breast cancer progression) increased the threshold at which reactive oxygen species (ROS) triggered phenotypic changes, suggesting that cellular response to ROS depends on the microenvironment. Theoretically, the GOC should be usable to also bring cells, like immune cells, in different concentrations to the cultured tissue.

 


Figure 1. Components of the Gradient-on-a-chip. (E) From left to right: top acrylic fixture with cell culture aperture, PET membrane (with microapertures) allowing access to the chemical gradient underneath and bringing the necessary stiffness to maintain the paper-based mixer inside the PDMS channels, paper microfluidic insert fitted to the PDMS channels to prevent leakage, PDMS microfluidic platform with laser ablated channels, and bottom acrylic fixture; the inset is a high-magnification image of the laser-machined microapertures on the PET membrane. (F) Final assembled device. Scale bars: (E) 1 cm, (F) 1.5 cm (From Chittiboyina/Rahimi et al., ACS Biomat Sci & Eng, 2018).
 

 
 

Figure 2. Gradient formation. Image of gradient formed with red and blue dyes injected in separate inlets one hour before. One inlet on the opposite side is used to remove the fluid. The gradient mixer is visible just below to the inlets, before the rectangular culture chamber. The device is assembled with screws. (From Chittiboyina/Rahimi et al., ACS Biomat Sci & Eng, 2018)
 

 


Sophie A. Lelièvre, D.V.M., Ph.D., LL.M.P.H.

Professor, Basic Medical Sciences

Coleader, Drug Delivery & Molecular Sensing, PUCCR

Scientific Director, 3D Cell Culture Core (3D3C) Facility

Coleader, International Breast Cancer & Nutriton (IBCN)


 

 

 


3D in Action


3D3C at the Experimental Biology meeting

 

3D3C was invited by Jessica Ellis (Purdue Nutrition Science), to organize a workshop under the tutelage of the American Society for Biochemistry and Molecular Biology during the Experimental Biology 2018 Annual Meeting in San Diego, CA. The 90-minute workshop was scheduled on April 22, 2018.

Workshop Title: The Art of the 3D Cell Culture--- from Organoids to Organs-on-a-Chip

Abstract submitted for the workshop:  When 3D cell culture is properly done the cells behave in a manner that allows their organization and function as in vivo. In the workshop participants will become familiar with the concepts used to place cells in an environment appropriate for their needs and for the study of interest, from simple contexts with one cell type, to complex tissue organization, with multiple cellular compartments.  They will participate in interactive observations/analyses of cells in 3D culture and build platforms for organs-on-a-chip from kits. Upon completing the workshop, participants will understand why there can be many cell culture models depending on the scientific queries and technical capabilities, with materials ranging from sophisticated polymers to paper; and they will be better prepared to choose a model convenient for their needs. 

Outcomes: The workshop was very well-received and the room was practically full, with approximately 150 attendees. The first part of the workshop consisted of an interactive one-hour presentation that stimulated many questions and discussion points, with the appreciation from the audience of the importance to think about tissue organization when working with 3D cell culture. For the 30-minute session with hands-on practices, 3D3C explored a new concept for the design of 3D cell culture that we call OCOrigami (Organ-on-a-Chip Origami), which consists of using paper-based kits to construct organ models. Here we used the examples of the lung and the liver. The attendees were very enthusiastic and actively involved in playing with paper folding and blowing into straws. Who ever thought that cell culture was not fun?!

 


3D in Focus

  

Cells interact mechanically with their microenvironment. On one hand, externally imposed forces on the matrix can impact cell behavior, on the other hand, the forces generated by cells in the microenvironment may propagate through the matrix and influence the mechanics of such extracellular milieu. However, the impact of cell-generated forces remains to be clearly understood. Notably, direct measurements of local matrix stiffness in three-dimensional (3D) contexts, at a cell level, were awaited. By probing the local micromechanics of cells, the authors of this issue’s highlighted studies were able to observe a remarkably far-reaching stiffening gradient generated in response to cell-generated forces in a variety of biopolymer matrices.
 

Yu Long Han, Pierre Ronceray, Guoqiang Xu, Andrea Malandrino, Roger D. Kamma, Martin Lenze, Chase P. Broederszf, and Ming Guo. Cell contraction induces long-ranged stress stiffening in the extracellular matrix, PNAS www.pnas.org/cgi/doi/10.1073/pnas.1722619115


3D3C summary of the article: Cells were cultured at a concentration that maintained cells isolated in extracellular matrix infused with latex beads. The local micromechanics of the matrix was probed using optical tweezers to pull the latex beads away from the cell at a constant speed. For human breast cancer MDA-MD-231 cells in collagen matrix, the matrix became substantially stiffer closer to the cells; a nonlinear stiffness gradient, as high as 50 Pa/μm, was observed. Inhibiting contractility of MDA-MB-231 cells with cytochalasin D (a disruptor of actin cytoskeleton) resulted in a strong attenuation of the cell-induced stiffening. For cells with weak contractility, like non-neoplastic human mammary epithelial MCF-10A cells, the stiffening of the surrounding matrix was found to be negligible. In other matrix systems, MDA-MB-231 cells in Matrigel and human umbilical vein endothelial cells (HUVECs) in fibrin gel, both cell types were capable of generating large, extended stiffness gradients along the contraction direction of cells.

The strong stiffness gradient occurring in the vicinity of a cell is proposed to be originated from the active forces that the cell exerts through the matrix. The matrix surrounding a strong contractile cell behaves as a network of ropes. In such case, the total contractile force exerted by the cell is conserved over a distance, and the decay of radial stress simply reflects force spreading over an increasing surface area. This buckling based mechanism for long-range stress transmission is supported by observations with confocal reflection microscopy of a larger amount of highly curved collagen filaments in the vicinity of a contractile cell, compared with the situation in which contraction is inhibited with cytochalasin D.

Comments from 3D3C: It has been demonstrated that extracellular matrix stiffness has profound impacts on cellular functions (as illustrated for instance in the gradient-on-a-chip work of the Lelièvre and Ziaie laboratories highlighted in this newsletter). In most studies, matrix stiffness is manipulated through matrix composition or applied external physical force. The cell-generated force on matrix stiffness, however, is rarely discussed, although it illustrates perfectly the general concept of dynamic reciprocity between cells and their extracellular matrix proposed by Mina J. Bissell in the early 80s (Bissell et al, J Theor BIol, 1982). Interestingly, as the highlighted studies confirmed the presence of a nonlinear far-reaching stiffness gradients generated by cells in a 3D matrix, we wonder how results would be changed if the cells were organized as a sheet of interacting cells like in a physiologically relevant epithelium. Surely, the strong cell-cell interactions would influence cell-ECM mutual influence. Another point to consider is how cells like fibroblasts, that strongly mechanically interact with the matrix, would sense epithelial cell-generated forces and mitigate such forces inside the matrix.
  


 


 3D in Publications


Recent publications on 3D culture (please click to access the list on our web page
https://nanohub.org/groups/3d3cfacility):

Review

The research articles and reviews are arranged in the following categories:
 

 

 


3D in Meetings

 

3rd International Conference on 3D Printing in Medicine
Date: 4th to 5th May 2018
Location: Halle, Germany
Website: http://3dprint-congress.com/
Contact person:congress@bb-mc.com
Organized by: boeld communication GmbH

 

2nd World Congress and Expo on Nanotechnology and Materials Science
Date: 25th to 27th June 2018
Location: Dubai, United Arab Emirates
Website: https://biocoreconferences.com/nanotechnology2018/index.php
Contact person: Hany
Organized by: Biocore



3D Tissue Models: Oncology 2018 
Date: 8th to 10th May 2018 
Location: Boston, MA, USA
Website: http://go.evvnt.com/194508-0?pid=80 
Contact person: Doug Fairbrother 
Organized by: Hanson Wade 
 

8th World Congress of Biomechanics, Dublin, Ireland 2018 
Date: 8th to 12th May 2018 
Location: Dublin, Ireland 
Website: http://go.event.com/173142-0 
Contact person: WCB 2018 Congress office 
Organized by: World Congress of Biomechanics, Dublin, Ireland 2018 



2nd World Congress and Expo on Nanotechnology and Materials Science 
Date: 25th to 27th June 2018                         
Location: Dubai, United Arab Emirates 
Website: https://biocoreconferences.com/nanotechnology2018/index.php  
Contact person: Hany 
Organized by: Biocore 
Deadline for abstracts/proposals: 25th May 2018 


EMBL Conference: Microfluidics 2018: New Technologies and Applications in Biology, Biochemistry and Single-Cell Analysis
Date: 15th to 17th July 2018 
Location: Heidelberg, Germany
Website: http://www.embl.de/training/events/2018/MCF18-01/index.html
Contact person: events@embl.de
Organized by: EMBL CCO
 
 
Gordon Research Conference — Signal Transduction by Engineered Extracellular Matrices
Date: 22nd to 27th July 2018 
Location: Andover, NH, USA
Website: http://www.grc.org//signal-transduction-by-engineered-extracellular-matrices-conference/2018/
Contact person: Sarah C. Heilshorn
Organized by: Gordon Research Conference
 

PREDiCT: 3D Models  
Date: 21st to 23rd August 2018 
Location: Boston, MA, USA
Website: http://go.evvnt.com/224119-2?pid=80  
Contact person: Doug Fairbrother 
Organized by: Hanson Wade 


4th International Conference on Bio-based Polymers and Composites
Date: 2nd to 6th September 2018
Location: Balatonfüred, Hungary
Website: http://www.bipoco2018.hu
Contact person: Dóra Tátraaljai
 
Bio-based polymers and their blends, composites. Natural polymers and their modification. Natural fiber reinforced composites. Other raw materials based on natural resources. Biodegradation and environmental issues

 

EMBO|EMBL Symposium: Organoids: Modelling Organ Development and Disease in 3D Culture
Date: 10th to 13th September 2018
Location: Heidelberg, Germany
Website: http://www.embo-embl-symposia.org/symposia/2018/EES18-08/index.html
Contact person: events@embl.de
Organized by: EMBL
 

The 4Bio Summit  
Date: 27th to 28th November 2018 
Location: Rotterdam, Netherlands 
Website: http://www.global-engage.com/event/4bio/?utm_source=4BIOEUMP2018 
Contact person: Jane Williams 
Organized by: Global Engage 
 
 
Gordon Research Conference — Biomaterials and Tissue Engineering
Date: 28th July to 2nd August 2019 
Location: Castelldefels, Spain
Website: http://www.grc.org//biomaterials-and-tissue-engineering-conference/2019/
 
Contact person: Jennifer L. West and Brendan A. Harley
Organized by: Gordon Research Conference