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R Kumar, E Handy, K Dutt, CA Nweke; Rotary Culture of Human Retinal Progenitors and Human Retinal Pigment Epithelial (RPE) Cells: Tissue Engineering . Invest. Ophthalmol. Vis. Sci. 2002;43(13):3628.
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Purpose: The purpose of this study was to test if retinal structures can be generated from transformed and non-transformed human retinal progenitors co-cultured with RPE as rotary aggregated culture using spheroid technology developed by others for dissociated primary cultures from animal eyes. Methods: Transformed and non-transformed human retinal progenitors (301, Ret1-4) were cultured alone or as a co-culture with a human RPE (D407) cell line for 1-14 days. Cell aggregates were evaluated by phase, scanning, and immunophenotyping and expression of neurotrophins was monitored by RT-PCR and Northern blot analysis. Results: RT-PCR and Northern blot analysis showed upregulation of bFGF, BDNF, TGFa and CNTF in rotary co-cultures of retinal progenitors and RPE. Western blot analysis showed increased expression of dopamine receptor D4 and PKCa. Conclusion: Non-transformed retinal cells co-cultured with RPE showed up regulation of neurotrophins and differentiation specific proteins PKCa and dopamine receptor D4. However, formation of laminated retina was not seen, possibly due to missing extracellular matrix molecules and/or growth factors. Future studies will explore the role of added extracellular matrix proteins in this culture system. CR: None Support: NASA Grants NCC-9-53 (KD); NAG-9-964 (KD); and NAG-9-1164 (64)
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