11 December 2015

Retinal nerve cells were grown in the laboratory

Johns Hopkins University researchers have developed an effective method of converting human stem cells into retinal ganglion cells – a type of nerve cells localized in the retina and providing the transmission of visual signals from the eye to the brain. The death and disruption of the functioning of these cells lead to loss of vision in diseases such as glaucoma and multiple sclerosis.

Using the CRISPR-Cas9 genetic editing method, the researchers embedded a red fluorescent protein gene into the DNA of human embryonic stem cells. Expression of this gene in cells should have been triggered only when another BRN3B (POU4F2) gene was expressed. The BRN3B gene is expressed in mature retinal ganglion cells, and its protein product is visible under a microscope in red light. Subsequently, this was used for fluorescence-activated selection of differentiated retinal ganglion cells from a mixed population.

A series of experiments on directed differentiation showed that the addition of a plant-derived compound forskolin on the first day of differentiation increased the efficiency of the transformation of stem cells into retinal ganglion cells. On the thirtieth day, clusters of fluorescent cells visible under the microscope appeared in the culture. The authors are very inspired by this result, as for the first time they were able to isolate retinal ganglion cells and study them in a pure population. They are currently searching for other genes that play important roles in the survival and functioning of ganglion cells.

Fluorescence microscopy: retinal ganglion cells on the 50th day of cultivation

According to the head of the study, Professor Donald Zack, the results obtained will not only help to better understand the biology of the optic nerve, but will also allow creating cellular models for the development of drugs. Moreover, they may eventually form the basis of cell transplant therapy methods for restoring vision in patients with glaucoma and multiple sclerosis.

It should also be noted that forskolin is widely available in the form of dietary supplements for weight loss and muscle building and is positioned as a herbal preparation for the treatment of various diseases, however, the authors warn that the effectiveness and safety of its use for the treatment or prevention of blindness or any other diseases have no scientific evidence.

Article by Valentin M. Sluch et al. The differentiation of human ESCs to retinal ganglion cells using a CRISPR engineered reporter cell line is published in the journal Scientific Reports.

Evgeniya Ryabtseva
Portal "Eternal youth" http://vechnayamolodost.ru based on Johns Hopkins University: Researchers Grow Retinal Nerve Cells in the Lab.  

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