Navigation Links
A 'clear' choice for clearing 3-D cell cultures
Date:9/3/2014

PROVIDENCE, R.I. [Brown University] Because Brown University biomedical engineering graduate student Molly Boutin needed to study how neural tissues grow from stem cells, she wanted to grow not just a cell culture, but a sphere-shaped one. Cells grow and interact more naturally in 3-D cultures than when they're confined to thin slides or dishes.

But the very advantage of a culture having thickness also poses a challenge: How to see all the cells and their connections all the way through the culture. It's a problem that confronts many biologists, physicians, bioengineers, drug developers- and others who also see 3-D cultures as a useful stage before moving to animal models.

"As I was imaging these tissues I was only able to get the outer layer or two of cells and that wasn't a very good representation of what was going on inside of the sphere," Boutin said.

There are inelegant ways to slice up an engineered 3-D tissue for imaging and then to reconstruct it, but a more tantalizing solution seemed likely to come from one of the chemical treatments invented in just the last few years to make tissues see-through. But which one, if any, would work with her scaffold-free engineered neural tissues? To find out, Boutin and adviser, Diane Hoffman-Kim, associate professor of medical science in the Department of Molecular Pharmacology, Physiology, and Biotechnology, decided to test the three simplest methods: ClearT2, SeeDB and Scale.

Their results which read like a Consumer Reports article for the lab bench set now appear online in the journal Tissue Engineering Part C: Methods.

For Boutin's critieria, the ClearT2 method turned out to be clear winner. For her little balls of neural tissue 100 millionths of a meter in diameter ClearT2 allowed her to see fluorescing cells at all depths of focus and, importantly, it did not change the size of the tissue. Scale made her "neural spheres" substantially larger, while SeeDB made them smaller and didn't improve clarity as much.

Maintaining the tissue culture size is important because Boutin wants to know the physical dimensions of growth in the culture, such as the axons that one neuron might extend to another.

While ClearT2 worked within 1.5 hours, Scale and SeeDB took three days, Boutin said.

She ran several further tests with ClearT2, including with other types of healthy and cancerous neural cells, and ClearT2 continued to perform well, even allowing her to image extracellular matrix.

In the paper Boutin and Hoffman-Kim acknowledge that the results with different methods may vary with different samples, but they expect that for many "scaffold-free" engineered 3-D neural tissues tissues that grow without added matrix supports ClearT2 should work well.

Knowing that could clear the way for many research projects with 3-D tissues.


'/>"/>

Contact: David Orenstein
david_orenstein@brown.edu
401-863-1862
Brown University
Source:Eurekalert

Related biology news :

1. IU biologists offer clearer picture of how protein machine systems tweak gene expression
2. New paper by Notre Dame researchers describes method for cleaning up nuclear waste
3. Report presents designs for study of cancer risks near US nuclear facilities
4. Dartmouth scientists track radioactive iodine from Japan nuclear reactor meltdown
5. Postpone the nuclear waste decision
6. Autologous bone marrow-derived mononuclear cell transplants can reduce diabetic amputations
7. Scientists core into Californias Clear Lake to explore past climate change
8. Researchers at GW receive federal funds to study the effect earthquakes have on nuclear reactors
9. Where to put nuclear waste?
10. Structure of RNAi complex now crystal clear
11. Europe clears the air
Post Your Comments:
*Name:
*Comment:
*Email:
(Date:2/28/2017)... 2017 News solutions for biometrics, bag drop ... ... 14 to 16 March, Materna will present its complete end-to-end ... travel is a real benefit for passengers. To accelerate the ... passenger touch point solutions to take passengers through the complete ...
(Date:2/24/2017)... Feb. 24, 2017  EyeLock LLC, a leader of iris-based ... iris biometric solution on the latest Qualcomm® Snapdragon™ ... Mobile World Congress 2017 (February 27 – ... Hall 3, Stand 3E10. The ... security platform—a combination of hardware, software and ...
(Date:2/16/2017)... , Feb. 16, 2017  Genos, a community ... that it has received Laboratory Accreditation from the ... is presented to laboratories that meet stringent requirements ... scientifically rigorous processes. "Genos is committed ... laboratory practices. We,re honored to be receiving CAP ...
Breaking Biology News(10 mins):
(Date:3/23/2017)... March 23, 2017 According to a report ... derivatives market is fragmented due to the presence of a large pool ... Thermo Fisher , and Sigma-Aldrich, compete with each other in ... collectively, held more than 76% of this market in 2016.  ... As of now, a ...
(Date:3/22/2017)... N.Y. , March 22, 2017 Regeneron Pharmaceuticals, ... the Regeneron Genetics Center (RGC), U.K. Biobank and GSK to ... the U.K. Biobank resource. The initiative will enable researchers to ... new medicines for a wide range of serious and life ... ...
(Date:3/22/2017)... March 22, 2017 Good Start Genetics, a ... eclipsed the 130 million covered lives mark through its ... Texas . With newly signed contracts ... to enjoy strong payor acceptance based on the quality ... and genetic counseling, its industry-leading customer care and support ...
(Date:3/22/2017)... MarketNewsUpdates.com News Commentary  ... The traditional ways to ... as of late due to the rise of the opioid ... dramatic impact on patient,s quality of life as Biotech and ... new forms of opioid formulations that prevent abuse. Biotech and ...
Breaking Biology Technology: