[BNC-all] 3D3C Newsletter - August 2016

Wed Sep 7 14:59:38 EDT 2016

Dear All,

Below please find the fifth 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 four sections in the newsletter are:
3D at Purdue – this section highlights 3D cell culture-based research activity at Purdue
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

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Volume 5, August 2016

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3D at Purdue

3D in Focus

3D in Publications

3D in Meetings

3D at Purdue

Multi-cellular, Multi-niche 3D Culture to Mimic the Brain Tumor Microenvironment

Glioblastoma (GBM), the most common adult primary brain tumor, has a mean life expectancy of about 15 months even with aggressive surgery, radiation and chemotherapy.  Both drug resistance and a highly invasive phenotype contribute to the lethality of this cancer.  Typically, by the time a neurosurgeon has an opportunity to intervene, cancer cells have already migrated throughout most of the brain. Like many cancer cells, GBM develops a dynamic relationship with the cell microenvironment to support tumor growth, migration and drug resistance. GBM cells find productive paths of migration along blood vessels and white matter tracts, and neighboring support cells, including normal astrocytes and pericytes, may support the survival and migration.

In order to tease out the role of the remodeled extracellular matrix and cell-cell interactions in GBM, we created 3D multicellular and multiniche cell culture models.  A tunable oligomeric collagen I (Geniphys) forms the base of the system to control the mechanical properties using a chemically well-defined, but natural material.  We find that GBM migration is most productive when the matrix is tuned to match the storage modulus range of the normal brain.  Hyaluronan (HA) is integrated with the collagen to mimic the chemical content of the brain, which is HA rich.  Basement membrane coated microfibers are embedded in the culture to structurally mimic blood vessels.  We observe that GBM cells alter their migration mode from a mesenchymal migration phenotype into a highly productive, collective migration when they encounter a pseudovessel.  Finally, human normal astrocytes and/or endothelial cells are also included as a coculture with the GBM cells in the matrix.  As predicted but never before shown, we see that astrocytes further enhance the migration of the GBM cells in 3D.  Both cell types alter the response to drugs, with different effects depending on the background characteristics of the starting tumor, which can vary widely from patient to patient.

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In the end, we hope to build a 3D tissue model of GBM that can better predict therapy outcomes and facilitate the design of multipronged strategies to finally beat glioblastoma in the clinic.

Jenna Rickus, PhD
Professor,  Agriculture and Biological Engineering, Biomedical Engineering

Herrera-Perez M. Voytik-Harbin SL, Rickus JL. “Extracellular Matrix Properties Regulate the Migratory Response of Glioblastoma Stem Cells in Three-Dimensional Culture.” Tissue Eng Part A. 2015 Oct;21(19-20):2572-82.
http://www.ncbi.nlm.nih.gov/pubmed/26161688

Funded with support from the Purdue Center for Cancer Research and the IUPUI Center for the Cure of Glioblastoma.

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3D in Focus

3D cell cultures should provide conditions for cells to differentiate into physiologically relevant tissue structures. Therefore, it is thought that 3D cell culture systems offer a better alternative for drug screening than conventional 2D culture methods. Improved understanding and characterization of 3D cell culture models are essential for further application in drug screening/discovery studies. One of the critical elements in 3D cell culture is the extracellular matrix since it greatly impacts the biological characteristic of cells in the culture system. This matter is addressed in the following article by Edmondson et al., which we will discuss in today’s newsletter. For those who attended, or plan to attend, future workshops on basics of 3D cell culture with gels, notably to induce tumor formation, this article is an example of how the type of matrix strongly influences cells in culture, and it highlights the importance of testing matrices for best mimicry of in vivo situations.

Edmondson R, Adcock AF, Yang L (2016) “Influence of Matrices on 3D-Cultured Prostate Cancer Cells' Drug Response and Expression of Drug-Action Associated Proteins.” PLoS ONE 11(6): e0158116.

In the study reported here, human prostate cancer LNCaP and DU145 cells were cultured in two biologically-derived matrices, Matrigel and Cultrex BME, and one synthetic matrix, the Alvetex scaffold. Both Matrigel and Cultrex BME are from Engelbreth-Holm-Swarm (EHS) sarcoma and rich in basement membrane type of extracellular matrix proteins including laminin, collagen IV, as well as growth factors such as epidermal growth factor and insulin-like growth factor. It seems that the main difference in this study for the choice of these two matrices was the protein concentration that was 8 – 12 mg/ml for Matrigel and 15 to 17 mg/ml for Cultrex BME. Alvetex is a synthetic, highly porous polystyrene scaffold. Note that EHS-derived gels provide natural extracellular matrix molecules to which cells react; with the polystyrene scaffold, the cells need to be able to synthesize and deposit the matrix molecules that they need for their survival and thrive, which is often the case with cancer cells.

DU145 and LNCaP cells in 3D culture proliferated at a slower rate than cells placed in 2D (standard) cultures. When comparing the effect of each matrix, DU145 cells proliferated at the highest rate in Alvedex, followed by Matrigel and Cultrex BME. Whereas, for LNCaP cells, the decreasing order for impact on proliferation was Matrigel, Alvedex and Cultrex BME.  Moreover, DU145 cells formed round-shaped spheroids in both Matrigel and Cultrex BME, but failed to form spheroid structures in Alvetex. Whereas, LNCaP cells formed mass-type spheroids in all three types of matrices.

The responses of tumor cells to anticancer drugs docetaxel and rapamycin in 3D culture were investigated. As shown in many other publications with different cell models and drugs, DU145 and LNCaP cells cultured in all three types of matrices were more resistant to docetaxel than those cultured in 2D. DU145 cells in Alvetex showed the highest resistance to docetaxel among the three types of matrices, while LNCaP cells showed similar response in all matrices. DU145 cells cultured in Matrigel and Cultrex BME were more sensitive to rapamycin that those cultured in Alvetex and in 2D (the latter two conditions triggered similar response to the drug). Unlike DU145 cells, LNCaP cells cultured in Matrigel and Cultrex BME were more resistant to rapamycin than those in Alvetex and 2D culture (the latter two conditions triggered similar response to the drug).

The authors speculated that the expressions of beta-III tubulin and EGFR correlated with the response of DU145 cells to docetaxel and rapamycin, respectively. The expression of beta-III tubulin followed a decreasing trend for cells cultured in Alvetex, Matrigel, BME, and 2D culture. Decreasing EGFR expression was observed in the following order, Alvetex, 2D culture, Matrigel, and BME.

Our conclusions:

Although this is a simple study and no suggestion regarding the role of the specific matrices in the biological response of cells can be proposed, the data from this study indicate that the type of matrix (even if in theory similar, as shown for Matrigel and Cultrex BME) will influence many cell features, such as morphology, proliferation, drug sensitivity, as well as gene expression. Furthermore, the impact of matrices is also cell line related.

There has been a lot of interest recently in applying 3D culture model in drug screening studies. The cells in 3D culture are cultured in a variety of conditions, as suggested by the present study, known to influence cellular characteristics and behavior. Therefore, further characterization and standardization of the 3D culture models will be critical for drug screening assays. There is a lot of work ahead of us.

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3D in Publications

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

Review<https://nanohub.org/groups/3d3cfacility/news#reviews>

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

Scaffold free/Scaffold
        Organ/Tissue/Cell       Others
Spheroids<https://nanohub.org/groups/3d3cfacility/news#spheroids>

Organoid<https://nanohub.org/groups/3d3cfacility/news#organoid>

Scaffold<https://nanohub.org/groups/3d3cfacility/news#scaffold>

Hydrogel<https://nanohub.org/groups/3d3cfacility/news#hydrogel>

Matrix<https://nanohub.org/groups/3d3cfacility/news#matrix>

Microfluidics<https://nanohub.org/groups/3d3cfacility/news#microfluidic>

Microfabrication<https://nanohub.org/groups/3d3cfacility/news#microfabrication>

Bone<https://nanohub.org/groups/3d3cfacility/news#bone>

Bone Marrow<https://nanohub.org/groups/3d3cfacility/news#bonemarrow>

Breast<https://nanohub.org/groups/3d3cfacility/news#breast>

Colon<https://nanohub.org/groups/3d3cfacility/news#colon>

Heart<https://nanohub.org/groups/3d3cfacility/news#heart>

Liver<https://nanohub.org/groups/3d3cfacility/news#liver>

Lung<https://nanohub.org/groups/3d3cfacility/news#lung>

Muscle<https://nanohub.org/groups/3d3cfacility/news#muscle>

Nerve<https://nanohub.org/groups/3d3cfacility/news#nerve>

Prostate<https://nanohub.org/groups/3d3cfacility/news#prostate>

Endothelial cells<https://nanohub.org/groups/3d3cfacility/news#endothelialcells>

Fibroblast<https://nanohub.org/groups/3d3cfacility/news#fibroblast>

Stem Cells<https://nanohub.org/groups/3d3cfacility/news#stemcells>

Stromal Cells<https://nanohub.org/groups/3d3cfacility/news#stromalcells>

Cancer/Tumor<https://nanohub.org/groups/3d3cfacility/news#cancer>

Screening<https://nanohub.org/groups/3d3cfacility/news#screening>

3D bioprinting<https://nanohub.org/groups/3d3cfacility/news#3dbioprinting>

Imaging<https://nanohub.org/groups/3d3cfacility/news#imaging>

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3D in Meetings

2nd EACR Conference on Goodbye Flat Biology: Models, Mechanisms and Microenvironment

Date: 2nd to 5th October 2016
Location: Berlin, Germany
Website: http://www.eacr.org/goodbyeflatbiology2016/index.php
Contact person: Roger Doxat-Pratt
The meeting should be of interest to all those who use cancer cell lines, patient-derived tissue samples or primary cultures in vitro for the study of tumour biology, bioengineering and biochemistry, drug target validation, etc.
Organized by: European Association for Cancer Research
Deadline for abstracts/proposals: 20th June 2016

14th Annual High-Content Analysis & 3D Screening

Date: 31st October to 2nd November 2016
Location: Cambridge, MA, USA
Website: http://www.highcontentanalysis.com/
Contact person: Jaime Hodges
Event will deliver the latest advancements in HCA applications and technologies, and the next steps for physiologically relevant complex models, ultra-high resolution and high-throughput imaging, and more advanced image analysis and data management.
Organized by: Cambridge Healthtech Institute

SPIE BiOS 2017 - Part of SPIE Photonics West 2017

Date: 28th January to 2nd February 2017
Location: San Francisco, CA, USA
Website: http://spie.org/SPIE-BiOS-conference
Contact person: Customer Service
BiOS 2017, part of SPIE Photonics West 2017, is the world’s largest biomedical optics and biophotonics conference. Topics range from biomedical optics, photonic diagnostic and therapeutic tools and systems, nano/biophotonics, and more!
Organized by: SPIE - The international society for optics and photonics
Deadline for abstracts/proposals: 18th July 2016

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