[BNC-all] 3D3C Newsletter - December 2016

Fri Jan 27 14:49:28 EST 2017

Dear All,

Below please find the seventh 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 7, December 2016

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3D at Purdue

3D in Focus

3D in Publications

3D in Meetings

3D at Purdue

Engineered Tissues
Using Electrohydrodynamic Based 3D Printing

The tumor microenvironment is a complex, multicomponent system that drives cancer cell processes such as proliferation, senescence, and metastasis. However, the roles of individual components of the tumor microenvironment can be difficult to evaluate. Factors such as the type and conformation of extracellular matrix (ECM) proteins, the spatial patterning of cells, and the mechanical properties of the local tissue environment all contribute to driving the transition of cancer cells from a proliferative state into a more invasive (metastatic) phenotype.

The Solorio lab is currently utilizing a 3D polymer fiber writing system that allows for the fabrication of hyper-porous polymer scaffolds with patterned fiber architectures. These structures consist of only 3% solid polymer material, and can be used to support complex networks of ECM proteins. The scaffolds maintain mechanical integrity under tensile and compressive forces from cells or convective fluid flow, and can be modified to provide surface functionality as well as control over mechanical properties. The low polymer mass minimizes interference of the scaffold with interactions between the cells and ECM proteins, while enhancing the mass transport of oxygen and nutrients. Utilizing a dynamic coating technique, protein fibrillogenesis is achieved within the open area of the polymeric constructs, resulting in a 3D network of ECM fibrils that fills the free volume of the scaffold. This dynamic coating technique has been used to create scaffolds with collagen, laminin, and fibronectin networks. These modular engineered microenvironments can be seeded with a variety of cell types, enabling investigation of cell-cell or cell-matrix interactions. As such, key elements of the native cancer microenvironment can be replicated. Furthermore, we have demonstrated the successful culture of primary human metastatic breast cancer cells obtained from clinical samples of pleural effusions and ascites, providing a potential avenue for chemotherapeutic drug screening as part of precision medicine strategies.

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Figure 1: Scaffolds are coated with fibronectin fibrils, filling the free volume of the scaffold (fibronectin green). Cells can then be seeded onto the scaffold and allowed to proliferate to create 3D engineered tissue (actin red, nucleus blue).

Luis Solorio
Assistant Professor
Weldon School of Biomedical Engineering

Selected Publications:

1. Xie F, Deng X, Kratzer D, Cheng K, Friedmann C, Qi S, Solorio L, Lahann J. Backbone-degradable polymers prepared by chemical vapor deposition, Angewandte Chemie International Edition 2017, 56: 203 –207

2. Solorio L, Exner AA. Effect of the subcutaneous environment on phase sensitive in situ forming implant drug release, degradation, and microstructure. Journal of Pharmaceutical Sciences, 2015; 104(12): 4322-28.

3. Solorio L, Perera RH, Wu H, Gangolli M, Silverman E, Hernandez C, Peiris PM, Broome, AM, Exner AA.  Nanobubble ultrasound contrast agents for enhanced delivery of thermal sensitizer to tumors undergoing radiofrequency ablation.  Pharmaceutical Research, 2014; 31(6): 1407-17.

4. Patel RB*, Solorio L*, Wu H, Krupka T, Exner AA.  Effect of injection site on in situ implant formation and drug release in vivo. Co-first author, Journal of Controlled Release, 2010 Nov 1;147(3): 350-8.

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3D in Focus

Microfluidic devices and spatial confinement -
Does fluid volume matter for cell differentiation?

Microfluidics-based cultures of cells, such as hepatocytes, are being developed as platforms for toxicology and tissue engineering applications in which cells need to be properly differentiated. For convenience, like to achieve high-throughput, cell cultures might not include a complete 3D environment (e.g., multiple layers of cells in the liver), and might only recapitulate the minimal structural and functional unit of the tissue, which renders the maintenance of differentiation more difficult. The microfluidics-based devices are popular because they permit the study of a very small population of cells and the precise control of the composition and continuous perfusion flow pattern of the medium bathing the cells. It is known that the medium flow is necessary to maintain the differentiation status of cells. However, in the device, the cells are arbitrarily cultured in a small volume of medium to minimize the use of reagents and fit in the microfluidic system; yet, we do not understand the impact of fluid volume on differentiation and what would happen if the fluid circulation were not an option for certain experiments. The objective of the article highlighted in this newsletter was to investigate the spatial confinement (i.e., small volume) effect on cellular differentiation, a usually less studied aspect of the tissue microenvironment.

Featured article: Haque A, Gheibi P, Gao Y, Foster E, Son KJ, You J, Stybayeva G, Patel D, Revzin A.  Cell biology is different in small volumes: endogenous signals shape phenotype of primary hepatocytes cultured in microfluidic channels. Sci Rep. 2016 Sep 29; 6:33980. doi: 10.1038/srep33980.

3D3C overview of the article: Primary rat hepatocytes were cultured on collagen I coated low-volume microfluidic channels; the microchamber was 1 μl in volume. Instead of continuous perfusion, the medium inside the microchambers was changed every 48 hours to allow time to witness the spatial confinement effect. The authors showed that hepatocytes cultured in the microchambers retained a differentiated hepatic phenotype for 21 days. The differentiation phenotype was illustrated by the presence of cellular polarity, high levels of E-cadherin and albumin synthesis.  The levels of E-cadherin expression, albumin synthesis, and cellular polarity were impaired when the local volume in the microchambers was increased from 1 μl to 30 μl.  Likewise, cells cultured in standard 12-well plates under identical conditions (same surface coating, seeding density and medium composition), but with a large amount of medium per well, were dedifferentiated after seven days.  Therefore, spatial confinement in microfluidic devices, in addition to or in place of medium flow, is able to maintain the differentiation status of cells for a long period of time. The authors further showed that the differentiated phenotype was maintained at least by accumulation of hepato-inductive growth factors, such as hepatocyte growth factor, and reduction in hepato-disruptive growth factors, such as transforming growth factor (TGF)-β1, in the microchambers.  According to the authors, spatial confinement, even though it has not been seen as an option, may be an important alternative in microfluidic devices to maintain the normal function of cells when continuous flow is not possible.

As it seems that spatial confinement in microchambers will help maintain the differentiation of cells, we are wondering if spatial confinement is also an important factor in vivo. We should keep in mind that the design of microfluidic devices ought to recapitulate as much as possible the conditions under which cells are in tissues. Hence, the volume of fluid or movement should mimic what exists in the tissue microenvironment. This step is currently difficult to implement because the spatial characteristics of the fluid surrounding individual cells or groups of cells in a given tissue is rarely known. The analysis of spatial confinement in real tissue is an element of the next frontier to help improve 3D cell culture devices.

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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

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

3D Cell Culture 2017

Date: 22nd to 23rd February 2017
Location: London, United Kingdom
Website: https://go.evvnt.com/69495-0
Contact person: Honey de Gracia
Organized by: SMi Group
Deadline for abstracts/proposals: 21st February 2017

Keystone Symposia - Engineered Cells and Tissues as Platforms for Discovery and Therapy

Date: 9th to12th March 2017
Location: Boston, Massachusetts, United States
Website: https://www.keystonesymposia.org/17K1
Contact: Phone: 1 800-253-0685;     Email: info at keystonesymposia.org<mailto:info at keystonesymposia.org>
Organized by: Keystone Symposium

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