Generation of Mouse Retinal Ganglion Cells from the Transdifferentiation of Male Germline Stem Cell Source
Refereed conference paper presented and published in conference proceedings


摘要Statement of the Problem: Glaucoma is characterized by retinal ganglion cell (RGC) degeneration and is the leading cause of blindness worldwide. However, current treatments such as eye drops or surgery are limited and do not target the loss of RGC. Regenerative therapy using embryonic stem cells (ESCs) holds a promising option but ethical concern hinders clinical applications on human subjects.
Methodology & Theoretical Orientation: We hypothesized that RGCs could be generated from mouse spermatogonial stem cells (SSCs). We collected SSCs from mouse testes and they were subsequently transdifferentiated to ESC-like cells (SSC-ESCs) through a non-viral and non-invasive approach. These cells then underwent retinal differentiation to differentiate towards retinal lineages. To enable efficient isolation of RGCs by fluorescence-activated cell sorting (FACS), we generated Brn3b-GFP reporter SSC-ESCs, with Brn3b being a specific RGC marker.
Findings: SSC-ESCs exhibited stem cell properties similar to ESCs in terms of morphology and pluripotency gene expression profile. SSC-ESCs differentiated towards retinal cell lineage and developed optic vesicles in 7-10 days in in vitro culture, while some cells generated neurites in 13-16 days. These neuron-like cells were positive for RGC-specific marker Brn3b and other neural markers, thus confirming their RGC identity. We also performed microelectrode array (MEA) and SSC-ESC-derived RGCs demonstrated response to light. GFP+ RGCs (8% of differentiated cells) were isolated by FACS and able to survive in subsequent in vitro culture.
Conclusion & Significance: We successfully generated functional RGCs from a male germline stem cell source. We are currently working on transplantation of the derived RGCs to glaucomatous retina to examine its regenerative effect.
著者Hoi Ching Ashley SUEN, Yan Tracy QIAN, Jinyue Jason LIAO, Chun Shui Alfred LUK, Wing Tung Annie LEE, Kai Shun Christopher LEUNG, Tsz Kin Michael NG, Wai Yee CHAN, Tin-Lap LEE
會議名稱Global Experts Meetings on Infectious Diseases & Stem Cell Biology

上次更新時間 2018-04-12 於 12:09