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Jishnu Saha, Elizabeth E Capowski, Divya Sinha, Jee Min, Joe Phillips, David M Gamm; Differential effects of early BMP4 exposure on optic vesicle production across multiple hiPSC lines.. Invest. Ophthalmol. Vis. Sci. 2016;57(12):1133.
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© 2017 Association for Research in Vision and Ophthalmology.
To test the effect of brief, early BMP4 treatment (Kuwahara et al. (2015) Nat Commun. 6:6286) on the production of 3-D optic vesicle-like structures (OVs) from multiple hiPSC lines.
hiPSC lines derived from 8 individuals were maintained on Matrigel in mTesR1 and differentiated to OVs according to our established stepwise protocol, which minimizes the use of exogenous factors (Meyer et al. (2011) Stem Cells 29: 1206; Phillips et al. (2012) IOVS 53:2007). On day 6 of differentiation, each line was treated with or without 55 ng/ml BMP4, followed by 1:1 media exchanges every 3 days until day 16, when neuroectodermal colonies were lifted from the culture plates. Efficiency of hiPSC-OV production was assessed by counting OVs and non-OV neurospheres at day 25.
In the absence of BMP4, one of the hiPSC lines routinely generated >50% OVs (high producer), 6 generated <10% OVs (low producer), and one never generated OVs or any other neuroectodermal derivative (non-producer). The low producing lines had the greatest responses to BMP4 treatment, significantly increasing OV production an average of 8.4 ± 2.1-fold (p = 0.0038) per 6 well starting plate of undifferentiated hiPSCs. The non-producing hiPSC line also responded to BMP4 treatment by generating neuroectoderm, but did not yield OVs. However, the high producing line showed no difference in OV production with or without BMP4 treatment.
BMP4 treatment is capable of significantly increasing the yield of OVs in low producing hiPSC lines. These results extend the previously published findings of Kuwahara et al. (2015) and demonstrate both the utility and the limitations of this culture modification.
This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.
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