Sequencing of Induced pluripotent stem cell derived retinal progenitor cells

Age-related macular degeneration (AMD) is a result of degeneration/damage of the retinal pigment epithelium (RPE) while retinitis pigmentosa (RP), an inherited early-onset disease, results from premature loss of photoreceptors. A promising therapeutic approach for both is the replacement of lost/damaged cells with human induced pluripotent stem cell (hiPSC)-derived retinal cells. We studied the chemistry of retinal progenitor cells derived from iPSC through our patented unified differentiation protocol with the aim to take the cells for clincal benefits to needly patients. RPE expressed tight junction proteins, showed pigmentation and ciliation, and secreted polarization-related factors vascular endothelial growth factor (VEGF) and pigment epithelium-derived factor (PEDF). PRP expressed neural retina proteins and cone and rod markers, and responded to KCl-induced polarization. Transcriptomic analysis demonstrated an increase in the expression of mature retinal tissue-specific genes coupled with concomitant downregulation of genes from undesired lineages. RPE transplantation rescued visual function in RCS rats shown via optokinetic tracking and photoreceptor rescue. PRP transplantation improved light perception in NOD.SCID-rd1 mice, and positive electroretinography signals indicated functional photoreceptor activity in the host's outer nuclear layer. Graft survival and integration were confirmed using immunohistochemistry, and no animals showed teratoma formation or any kind of ectopic growth in the eye. We analysed Induced pluripotent stem cell and their derivatives -RPE and photoreceptor progenitor (PRP) cells at different timepoints. Following are the samples included in the study (1) Induced pluripotent stem cell (2) Retinal progenitors- obtained by dual SMAD and WNT inhibition from iPSC (3) RPE progenitors - obtained by removing the rosettes and culturing the epithelial pockets (4) Retinal Pigment Epithelium - pigmented RPE progenitors (5) Photoreceptor precursors - Rosette structures obtained in retinal progenitors enzymatically selected and expanded in culture. De novo-generated RPE and PRP were analyzed by Next generation sequencing.