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Generation of Retinal Organoids from Healthy and Retinal Disease-Specific Human-Induced Pluripotent Stem Cells
Published on: December 9, 2022
Molecular characterization of human retinal progenitor cells
Scott Schmitt1, Unber Aftab, Caihui Jiang
1Schepens Eye Research Institute, Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts 02114, USA.
Investigative Ophthalmology & Visual Science
|June 26, 2009
Summary
Fetal human retinal progenitor cells (hRPCs) maintain progenitor characteristics up to passage 6. Co-culture with mouse retina alters gene expression and confirms cell functionality.
Area of Science:
- Ophthalmology
- Developmental Biology
- Stem Cell Research
Background:
- Human retinal progenitor cells (hRPCs) are crucial for retinal development.
- Understanding hRPC behavior in vitro is vital for regenerative medicine strategies.
Purpose of the Study:
- To characterize the molecular profile of in vitro expanded fetal hRPCs.
- To investigate the impact of co-culture with mouse retina on hRPCs.
- To assess the functional capacity of hRPCs via calcium dynamics.
Main Methods:
- Isolation and in vitro expansion of fetal hRPCs (14-18 weeks GA).
- Analysis using immunoblot, microarray, and immunocytochemistry (ICC).
- Functional assessment via neurotransmitter-stimulated calcium imaging.
Main Results:
- hRPCs maintained progenitor markers (Nestin, PAX6) up to passage 6.
- Microarray confirmed expression of key retinal developmental genes.
- Co-culture significantly upregulated MKI67, PAX6, SOX2, GNL3, SIX3, and RHO.
- Calcium imaging demonstrated functional responses to neurotransmitters.
Conclusions:
- Fetal hRPCs retain progenitor identity through passage 6.
- These cells are functionally responsive to neurotransmitters.
- Co-culture with mouse retina modulates hRPC gene expression profiles.
