Stem cells grown into brain cells on a new 3D scaffold of tiny protein fragments

An MIT engineer and Italian colleagues will report the invention-which may one day replace the ubiquitous Petri dish for growing cells-in PLoS ONE.

Shuguang Zhang, associate director of MIT's Center for Biomedical Engineering, is a pioneer in coaxing tiny fragments of amino acids called self-assembling peptides to organize themselves into useful structures. Working with visiting graduate student Fabrizio Gelain from Milan, Zhang created a designer scaffold from a network of protein nanofibers, each 5,000 times thinner than a human hair and containing pores up to 20,000 times smaller than the eye of a needle.

The researchers were able to grow a healthy colony of adult mouse stem cells on the three-dimensional scaffold without the drawbacks of two-dimensional systems.

In addition to helping researchers get a more accurate picture of how cells grow and behave in the body, the new synthetic structure can provide a more conducive microenvironment for tissue cell cultures and tissues used in regenerative medicine, such as skin grafts or neurons to replace brain cells lost to injury or disease.

The scaffold itself can be transplanted directly into the body with no ill effects.

"The time has come to move on from two-dimensional dishes to culture systems that better represent the natural context of cells in tissues and organs," said Zhang, whose coauthors on the paper, in addition to Gelain, are from institutes and medical schools in Milan, Italy.

Biomedical researchers have become increasingly aware of the limitations of growing living cells in coated, two-dimensional Petri dishes and glass slides.

In the body, cells are attached to and supported by the cells, other structures and proteins around them. A cell's normal environment is a complex network of tiny fibers, gaps and pores through which oxygen, hormones and nutrients are delivered and waste products filtered away. Cells move within their natural environments in response to chemical signals or other stimuli.

Researchers are aware that cells on flat surfaces have skewed metabolisms, gene expression and growing patterns. But the only choices have been glass labware and a product called Matrigel, a gelatinous protein mixture secreted by mouse tumor cells. While Matrigel does resemble a complex extracellular environment, it also contains growth factors and unknown proteins that limit its desirability for experiments requiring precise conditions.

"Synthetic biopolymer microfiber scaffolds have been studied for more than 30 years to mimic a living 3D microenvironment, but concerns exist about their degradation products and chemicals," the authors wrote in the paper.

Other synthetic polymer biomaterials are simply too big. Getting cells to grow on them is like forcing spiders to build webs on skyscraper girders. Zhang's nanofiber scaffold, around 1,000 times smaller than the existing systems, is much closer in size to the extracellular matrices that living cells manufacture themselves.

http://www.mit.edu

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