[BNC-all] 3D3C Newsletter - October 2016

[Kwok, Tim]

Dear All,

Below please find the sixth 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 6, October 2016



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3D at Purdue

3D in Focus

3D in Publications

3D in Meetings



3D at Purdue


New Thermogels for Culturing Cells in 3D Structures

Kinam Park and Bumsoo Han

Culturing cells in 3D structures provides many advantages over 2D models. Cells in 3D structures often behave differently from those in 2D cultures.  For example, cancer cells organized in 3D spheroids are often more resistant to anticancer drugs than cells in 2D cultures. Several types of commercially available products allow researchers to produce multicellular spheroids.  The products are classified into (i) extracellular matrices based on cellular extracts, (ii) natural polymers, and (iii) cell culture plates with non-cell adherent surface or with no surface for the cells to attach.  Each method has its own advantages and limitations.

Recently, we developed simple polymeric systems that allow cells to form 3D structures.  The polymers are named “3DCellMaker” for its ability to culture various types of cells and cell mixtures in 3D (Fig. 1). 3DCellMakers are inverse themosensitive gels (thermogels), i.e., they remain in liquid form at room temperature or lower and become a gel at 37 °C. These thermogels are poloxamers-poly(ester-urethane) copolymers or stearate-modified methyl cellulose.  For the culture process, the cells can be placed on top of the formed gels, or they can be mixed with the thermogels at low temperature before incubating at 37 °C.  The simplicity, and low cost, of using 3DCellMaker allows culturing of various cell types in 3D for high throughput screening.  The synthetic nature of the 3DCellMaker makes it easy to control the gel’s properties.  The 3DCellMaker also allows culture of cells with defined additives, under serum-free conditions (Fig. 2).  3DCellMaker thermogels are available for free for researchers at Purdue University.


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Fig. 1. Co-culture of MCF-7 breast cancer cells and CCD-1068SK fibroblasts in a 3DCellMaker for five days. (Scale bar is 100 •m).












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Fig. 2. Culture of T4-2 breast cancer cells after two days in a 3DCellMaker under a serum-free condition (20X objective; courtey picture from the Lelièvre laboratory).














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3D in Focus

Angiogenesis is the process for new blood vessel formation through sprouting of endothelial cells from existing vessels. This process is regulated by biochemical and mechanical pathways. However, the effects of mechanical stimuli remain poorly appreciated and understood. Recent studies using microfabricated 3D culture systems have highlighted critical roles for flow-induced mechanical stimuli in angiogenesis. In this newsletter we are highlighting the work of Kim and colleagues on the role of mechanical stimuli in angiogenic sprouting using microfluidics.

Sudong Kim, Minhwan Chung, Jungho Ahn, Somin Leeb and Noo Li Jeon “Interstitial flow regulates the angiogenic response and phenotype of endothelial cells in a 3D culture model” Lab Chip, 2016, 16, 4189 - 4199

The authors have designed a microfluidic system with six parallel microchannels for the investigation of individual or combined effects of biochemical and mechanical factors during the initiation and outgrowth of angiogenic sprouts within the extracellular matrix. One of the two central channels is loaded with a fibrin matrix in which endothelial cells (ECs) are embedded (channel N); the other central channel contains an acellular fibrin matrix (channel S). The two outermost channels are used for the culture of stromal fibroblasts in the presence of fibrin. Fluidic channels located in-between the central and outermost channels are filled with culture medium with a flow at physiological level 0.1 – 4 μm S-1. The intent is for the soluble proangiogenic factors secreted by the fibroblasts to act on the ECs via the fluidic channels.

The initial phase of angiogenesis includes angiogenic sprout, vacuole formation within endothelial cells, and formation of a vascular network. Under static conditions in the device, angiogenic sprout extended from the cytoplasm of ECs (channel N), and these cells initiated vacuole formation that progressively developed into complex vascular networks. The sprouts extended first near the interface of channels N and S, and further into adjacent channel S where the ECs organized multicellular structures characterized by abundant actin-rich filopodia at their edges distal from the N channels thus, morphologically replicating the sprout tip of endothelial cells in vivo.

Moreover, the interstitial flow in the S to N direction in the fluidic channels promoted the formation of sprouts with a significantly greater number of filopodia projections compared to those formed under static conditions. In contrast, interstitial flow in the N to S direction completely inhibited the angiogenic sprouting. Unlike angiogenic sprouts, vacuole formation and the development of a vascular structure in channel N were not affected by the direction of the flow. Additional experiments confirmed that the interstitial flow controlled the initiation and sustained the growth of angiogenic sprouting.  Reversing the direction of the interstitial flow to S-N abolished the suppressor effect of N-S flow on sprouting.  On the other hand, reversing the flow direction from S-N to N-S had no effect on the existing vascular structure, but it induced less filopodia projections and the retraction of the forefront of the vasculature.

The model was subsequently examined for its capability to evaluate endothelial responses (i.e., sprout initiation, formation of vacuoles, vascular network formation) to extrinsic proangiogenic factors, such as VEGF, as well as anti-angiogenic compounds such as axitinib.

Angiogenesis is fundamental to many physiologic and pathologic processes such as cancer, ischemic diseases, and chronic inflammation. By acting as a proangiogenic stimulant, the interstitial flow in blood vessels can modulate the endothelial responses triggered by the biochemical stimuli such as VEGF.  The authors propose that the device can help elucidate the relevant mechanisms and thus, may lead to the development of effective therapeutics for diseases associated with angiogenesis.

The model designed here is interesting because of the addition of fibroblasts compared to the usual stand-alone endothelial cells to study angiogenesis within a microfluidics platform. The fibroblasts provide the angiogenic factors, making the model more physiologically relevant.




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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


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


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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