Nav: Home

Researchers grow retinal nerve cells in the lab

November 30, 2015

Johns Hopkins researchers have developed a method to efficiently turn human stem cells into retinal ganglion cells, the type of nerve cells located within the retina that transmit visual signals from the eye to the brain. Death and dysfunction of these cells cause vision loss in conditions like glaucoma and multiple sclerosis.

"Our work could lead not only to a better understanding of the biology of the optic nerve, but also to a cell-based human model that could be used to discover drugs that stop or treat blinding conditions," says study leader Donald Zack, M.D., Ph.D., the Guerrieri Family Professor of Ophthalmology at the Johns Hopkins University School of Medicine. "And, eventually it could lead to the development of cell transplant therapies that restore vision in patients with glaucoma and MS."

The laboratory process, described in the journal Scientific Reports, entails genetically modifying a line of human embryonic stem cells to become fluorescent upon their differentiation to retinal ganglion cells, and then using that cell line for development of new differentiation methods and characterization of the resulting cells.

Using a genome editing laboratory tool called CRISPR-Cas9, investigators inserted a fluorescent protein gene into the stem cells' DNA. This red fluorescent protein would be expressed only if another gene was also expressed, a gene named BRN3B (POU4F2). BRN3B is expressed by mature retinal ganglion cells, so once a cell differentiated into a retinal ganglion cell, it would appear red under a microscope.

Next, they used a technique called fluorescence-activated cell sorting to separate out the newly differentiated retinal ganglion cells from a mixture of different cells into a highly purified cell population for study. The cells showed biological and physical properties seen in retinal ganglion cells produced naturally, says Zack.

Researchers also found that adding a naturally occurring plant chemical called forskolin on the first day of the process helped improve the cells' efficiency of becoming retinal ganglion cells. The researchers caution that forskolin, which is also widely available as a weight loss and muscle building supplement and is touted as an herbal treatment for a variety of disorders, is not scientifically proven safe or effective for treatment or prevention of blindness or any other disorder.

"By the 30th day of culture, there were obvious clumps of fluorescent cells visible under the microscope," says lead author Valentin Sluch, Ph.D., a former Johns Hopkins biochemistry, cellular and molecular biology student and now a postdoctoral scholar working at Novartis, a pharmaceutical company. Sluch completed this research at Johns Hopkins before transitioning to Novartis.

"I was very excited when it first worked," Sluch says. "I just jumped up from the microscope and ran [to get a colleague]. It seems we can now isolate the cells and study them in a pure culture, which is something that wasn't possible before."

"We really see this as just the beginning," adds Zack. In follow-up studies using CRISPR, his lab is looking to find other genes that are important for ganglion cell survival and function. "We hope that these cells can eventually lead to new treatments for glaucoma and other forms of optic nerve disease."

To use these cells to develop new treatments for MS, Zack is working with Peter Calabresi, M.D., professor of neurology and director of the Johns Hopkins Multiple Sclerosis Center.
-end-
This work was completed through the Wilmer Eye Institute's Glaucoma Center of Excellence and its Stem Cell Ocular Regenerative Medicine Center, of which Zack is co-director.

Study co-authors were Chung-ha Davis, Vinod Ranganathan, Kellin Krick, Russ Martin, Cynthia Berlinicke, Nicholas Marsh-Armstrong and Hai-Quan Mao of Johns Hopkins; and Jeffrey Diamond and Justin Kerr of the National Institute of Neurological Disorders and Stroke.

This work was supported by grants from the Maryland Stem Cell Research Fund under grant number 2014-MSCRFI-0774; the National Institutes of Health under grant numbers T32-90040730, 1RO1EY02268001, 5T32EY007143, 5P30EY001765 and R01EY023754; and unrestricted funds from Research to Prevent Blindness Inc., the Guerreri Family Foundation, and Mr. and Mrs. Robert and Clarice Smith.

Johns Hopkins Medicine

Related Stem Cells Articles:

Computer simulations visualize how DNA is recognized to convert cells into stem cells
Researchers of the Hubrecht Institute (KNAW - The Netherlands) and the Max Planck Institute in Münster (Germany) have revealed how an essential protein helps to activate genomic DNA during the conversion of regular adult human cells into stem cells.
First events in stem cells becoming specialized cells needed for organ development
Cell biologists at the University of Toronto shed light on the very first step stem cells go through to turn into the specialized cells that make up organs.
Surprising research result: All immature cells can develop into stem cells
New sensational study conducted at the University of Copenhagen disproves traditional knowledge of stem cell development.
The development of brain stem cells into new nerve cells and why this can lead to cancer
Stem cells are true Jacks-of-all-trades of our bodies, as they can turn into the many different cell types of all organs.
Healthy blood stem cells have as many DNA mutations as leukemic cells
Researchers from the Princess Máxima Center for Pediatric Oncology have shown that the number of mutations in healthy and leukemic blood stem cells does not differ.
New method grows brain cells from stem cells quickly and efficiently
Researchers at Lund University in Sweden have developed a faster method to generate functional brain cells, called astrocytes, from embryonic stem cells.
NUS researchers confine mature cells to turn them into stem cells
Recent research led by Professor G.V. Shivashankar of the Mechanobiology Institute at the National University of Singapore and the FIRC Institute of Molecular Oncology in Italy, has revealed that mature cells can be reprogrammed into re-deployable stem cells without direct genetic modification -- by confining them to a defined geometric space for an extended period of time.
Researchers develop a new method for turning skin cells into pluripotent stem cells
Researchers at the University of Helsinki, Finland, and Karolinska Institutet, Sweden, have for the first time succeeded in converting human skin cells into pluripotent stem cells by activating the cell's own genes.
In mice, stem cells seem to work in fighting obesity! What about stem cells in humans?
This release aims to summarize the available literature in regard to the effect of Mesenchymal Stem Cells transplantation on obesity and related comorbidities from the animal model.
TSRI researchers identify gene responsible for mesenchymal stem cells' stem-ness'
Researchers at The Scripps Research Institute recently published a study in the journal Cell Death and Differentiation identifying factors crucial to mesenchymal stem cell differentiation, providing insight into how these cells should be studied for clinical purposes.
More Stem Cells News and Stem Cells Current Events

Trending Science News

Current Coronavirus (COVID-19) News

Top Science Podcasts

We have hand picked the top science podcasts of 2020.
Now Playing: TED Radio Hour

Listen Again: Reinvention
Change is hard, but it's also an opportunity to discover and reimagine what you thought you knew. From our economy, to music, to even ourselves–this hour TED speakers explore the power of reinvention. Guests include OK Go lead singer Damian Kulash Jr., former college gymnastics coach Valorie Kondos Field, Stockton Mayor Michael Tubbs, and entrepreneur Nick Hanauer.
Now Playing: Science for the People

#562 Superbug to Bedside
By now we're all good and scared about antibiotic resistance, one of the many things coming to get us all. But there's good news, sort of. News antibiotics are coming out! How do they get tested? What does that kind of a trial look like and how does it happen? Host Bethany Brookeshire talks with Matt McCarthy, author of "Superbugs: The Race to Stop an Epidemic", about the ins and outs of testing a new antibiotic in the hospital.
Now Playing: Radiolab

Dispatch 6: Strange Times
Covid has disrupted the most basic routines of our days and nights. But in the middle of a conversation about how to fight the virus, we find a place impervious to the stalled plans and frenetic demands of the outside world. It's a very different kind of front line, where urgent work means moving slow, and time is marked out in tiny pre-planned steps. Then, on a walk through the woods, we consider how the tempo of our lives affects our minds and discover how the beats of biology shape our bodies. This episode was produced with help from Molly Webster and Tracie Hunte. Support Radiolab today at Radiolab.org/donate.