Evaluation of Different Culture Media to Differentiate Human Embryonic Stem Cells Toward Eye Field Specification

Background and Aim: Degeneration of retinal pigment epithelium (RPE)or photoreceptors is one of the common causes of blindness around theworld. Most of recently replacement therapies are focused on humanembryonic stem cells (hESCs) derived RPE or photoreceptor. Severaldifferentiation protocols are under consideration for the generation ofhighly efficient retinal cells. During differentiation of hPSCs to rostral cellfate, one of the most important components is the cell culture media.Methods: In this study, we compared two widely used basic media,GMEM, and DMEM/F12, in order to assess the effects of culture mediainducing the efficient generation of rostral differentiation. The hESCswere cultured on Matrigel-coated dishes in the presence of two differentmedia which were supplemented with the same components including10% KSR, 2 mM L-glutamine and 0.1 mM NEAA. The morphologicalchanges were followed for one month. Also, the expression of specificeye filed markers at the RNA level was characterized.Results: Routinely, the hESCs were maintained in DMEM/F12supplemented by 20% KSR. To whether GMEM medium could inducerostral differentiation rather than DMEM/F12, in addition to declining KSRconcentration, in some groups the medium was changed to GMEM. Theresults showed that the morphology of hESCs was changed significantlyin the presence of GMEM. Quantification of the pigmented area as arostral differentiation criterion as well as neural tube-like structures waseffectively higher in GMEM medium when the results were comparedto DMEM/F12 as the control group. The assessment of eye field markersrevealed the efficiently inductive effect of GMEM medium to rostraldifferentiation of hESCs in compare to DMEM/F12.Conclusion: In conclusion, although DMEM/F12 medium is widely usedin many in vitro research studies such as proliferation and differentiationof hESCs, our results demonstrated GMEM would be a better replacementmedium for rostral differentiation especially in vitro eye field specificationstudies.

کلیدواژه ها:

Culture medium ، Human embryonic stem cell ، Cell differentiation ، Eye field