• Retinal Pigment Epithelium Differentiation

Retinal Pigment Epithelium (RPE) Differentiation

Human iPSC-derived retinal pigment epithelium (RPE) provides a physiologically relevant cell line model to understand basic ocular biology and ocular diseases. ASC’s iPSC-derived RPE protocol provides high quality RPEs with expression of lineage-committed markers and functional phenotype such as phagocytosis, and by providing an unlimited source of these cells.

  • RPE-like cells with typical cobblestone morphology and pigmentation
  • High quality and purity cells (60-80% purity) expressing RPE-specific markers Bestrophin 1 (BEST1)/ RPE65
  • iPSC reprogrammed from fibroblast/ PBMC/ CD34+ cord blood cells
  • From healthy/ disease/ engineered iPSCs
  • Optional! Control lines differentiated from “master” iPSC lines available
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Technical Details

Traditionally, in vitro modeling of retinal pigment epithelium (RPE) cells to understand ocular biology and ocular diseases has used primary RPE from donor tissues or immortalized cell lines. While the primary RPEs exhibit in vivo-like characteristics, they are difficult to source in large quantities from same donor. On the other hand, the immortalized cell lines like hTERT-RPE have overcome sourcing issues but do not shown the same gene expression or functional characteristics of in vivo RPEs are not truly representative.

iPSC-derived RPEs represent the best of both worlds: as primary cells they show in vivo-like genetic and functional characteristics and with the ability to generate large quantities of isogenic cell lines from the same parental iPSC line. In fact, transplantation of autologous iPSC-derived RPEs have been successful in rescuing vision in animal models and are being tested in patients in one of the first clinical trials in regenerative medicine. ASC provides custom service for iPSC differentiation to RPEs using optimized, efficient protocols:

  • RPE-like cells with typical cobblestone morphology and pigmentation
  • High quality and purity cells (60-80% purity) expressing RPE-specific markers Bestrophin 1 (BEST1)/ RPE65
  • From iPSCs derived using different reprogramming strategies from fibroblast/ PBMC/ CD34+ cord blood cells
  • From healthy/ disease/ engineered iPSCs
  • Well-characterized “Master” control, human iPSC lines available for a ready start to your differentiation projects as well as control lines for your disease modeling experiments

Optional! RT-PCR, Western Blot, proliferation assays, photoreceptor outer segment (POS) phagocytosis and other functional assays available upon request.

Applications:

  • Disease modeling such an age-related macular degeneration (AMD), diabetic retinopathy
  • Preclinical regenerative medicine research (adoptive transfer)
  • Drug testing and toxicity screening
  • High throughput drug target discovery

Timeline = 16-20 weeks

Deliverables

  • Cryopreserved iPSC-derived RPE: 60-80% BEST1+/RPE65+ cells
  • Antibody staining images and/or flow cytometry data for the cell type specific markers
  • Recovery Test, Mycoplasma Test
  • Sub-culture protocol and media (optional)
  • Other markers (available upon request)

Other related services:

 

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