Pre-adhesion promotes human stem cell-derived retinal ganglion cell survival and axon growth after transplant

Background Stem cell-derived human retinal ganglion cell (hRGC) transplant is a promising approach for vision restoration in optic neuropathies including glaucoma, but limited donor survival and axon regeneration remain major challenges. Methods We tested whether preconditioning by pre-attachment or host modulation improves organoid-derived hRGCs’ integration after intravitreal transplant. Cross-over transplant assessed the contribution of donor non-RGC populations. Netrin was delivered retrobulbarly to guide axon based on single-cell RNA sequencing (scRNA-seq) findings. Results The pre-attached donor cells exhibited significant survival and axon regeneration on the host retina (N=10-15 per group), with axons extending into the host optic nerve. Co-transplanted non-RGC populations further promotes survival and regeneration (N=5-10 per group). scRNA-seq revealed increased hRGC maturation after pre-attachment and transplant, including upregulation of axon guidance receptors. Exogenous chemoattractant netrin delivery significantly increased axon extension into the optic nerve head (ONH) (N=14-18 per group). Conclusion Pre-attachment and netrin guidance improved hRGCs integration after transplant, supporting therapeutic strategies for visual pathway restoration.