Generation Of Hair-cell Like Cells By Differentiation Of Human Umbilical Cord Mesenchymal Stem Cells In Vitro

Hearing loss is the most common form of sensory impairment of humans in industrialized nations, affecting people of any age and manifesting in various forms that rang in severity from modest difficulty with speech comprehension to profound deafness. Almost 360 million people around the world suffer from moderate to profound hearing loss. And the most prevalent of hearing impairment is sensorineural hearing loss(SNHL), which is caused by a range of genetic and environmental factors that degenerate and damage the inner ear hair cells and the spiral ganglion neurons. In adult mammals, auditory hair cells are terminally and well differentiated unique cells which have almost lost their regenerative potential after development. So if we can find a way to generate inner ear sensory epithelium from stem cells of other organs and restore the lost hair cells and spiral ganglion neurons by cell transplantation, there will be a hope for the curing of SNHL. Human umbilical cord mesenchymal stem cells(h UC-MSCs) share characteristics with other types of stem cells, such as the ability of sef-proliferation and multiple differentiation potential. Moreover, h UC-MSCs have a plentiful and inexpensive source, and is easy to get. They are lack of ethical and legal controversies, low immunogenicity and have the capability of immune-regulation. HUC-MSCs can be induced to form bone, cartilage, neural stem cells(NSC), liver like cells and amenable to biomedical engineering applications and cell transplantation therapy. In this study, we evaluated the potential h UC-MSCs as a possible source for regenerating inner ear hair cells in vitro. And we hope to explore a possible type of seed cells for the therapy of SNHL by inner ear cell transplantation. ObjectiveTo study the potential and protocol of human umbilical cord mesenchymal stem cells(h UC-MSCs) to differentiate towards hair-cell like cells in vitro. Methods1?Isolate human umbilical cord mesenchymal stem cells(h UC-MSCs) from Wharton's jelly by the regular way of our lab. Generate neural stem cells(NSC) from P3-P5 h UC-MSCs in DMEM/F12+N2+B27+EGF+b FGF medium within non-adherent cultures about 3-5 days. Use immunofluorescence to detect the expression of Nestin in induced cells.2?Treat NSCs derived from h UC-MSCs with DMEM/F12+N2+B27+EGF+ATRA in gelatin coated flask about 2-4 weeks to obtain inner ear hair cells. Use immunofluorescence and q RT-PCR to detect the expression of Math1?Myosin VIIa?Brn3c in the induced hair cells. Results1?HUC-MSCs trans-differentiated into neural sphere like cells and the induced cells expressed Nestin.2?Hair cell like cells were obtained from NSCs derived from h UC-MSCs and expressed Math1?Myosin VIIa?Brn3c. ConclusionhUC-MSCs can cross embryonic germ layers and trans-differentiate into neural stem cells, then subsequently to hair-cell like cells, thus may introducing a new potential tissue engineering and cell transplantation approach for the treatment of hearing loss.