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Deepti Singh, Pierre Colombe, Michael J Young; Exploring effect of culture condition on human retinal progenitor cells. Invest. Ophthalmol. Vis. Sci. 2019;60(9):3342.
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Using stem and progenitor cells to treat retinal disorders holds great promise and using defined conditions to maintain native phenotype is of clinical importance. We have cultured human retinal progenitor cells (hRPC) in different conditions such as normoxia and with knockout serum replacement (KOSR) to evaluate its effect on these cells. The purpose of this study was to identify if KOSR can replace commonly used growth factors in media
Human retinal progenitor cells were cultured in presence of defined medium along with growth factor that acted as control. In the experimental setup growth factors were replaced with knockout serum. Cells expression were analyzed with MACSquant using different antibody to check for stemness marker (OCT4 and Cmyc), proliferation (Ki67), Rods (Rhodopsin), Retinal maturation marker (Recoverin) and Retinal progenitor marker (Pax6).
Our results indicate cells cultured with KOSR in normal and hypoxia conditions do not show significant difference in viability. However cells with KOSR in normal oxygen condition expressed significantly higher stemness markers such as C-myc and OCT4 (31.20% and 13.44 % respectively) in comparison to hRPC cultured in KOSR at hypoxia (12.07% and 4.05%). Furthermore, markers for commitment such as rhodopsin were significantly decreased in the KOSR supplemented culture when compared to control. Similarly, normoxia has significant effect on the retinal maturation marker (Recoverin) that was found to decrease in comparision with control and KOSR in hypoxia.
KOSR is known to improve proliferation and maintain stemness of embryonic cells and our experiments suggest that hRPCs diverge into two distinct populations in the presence of KOSR. These results will help understand the effect of culture conditions and standardize differentiation protocols for clinical applications.
This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.
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