Abstract
Purpose :
To investigate the differentiation competence of human adipose tissue-derived stem cells (hADSCs) toward retinal lineage by inhibiting BMP and Wnt signaling.
Methods :
hADSCs were divided into induction group and negative control group (NC). hADSCs of induction group were induced toward retinal lineage with a 2-step induction procedure using media containing Noggin and Dkk1, antagonist of BMP signaling and Wnt/β-catenin signaling respectively. Step 1: hADSCs were induced into neurospheres on ultra-low attachment culture plates for 3 days. Step 2: Neurospheres were transferred onto matrigel-coated culture plates and induced to differentiate for up to 24 day. hADSCs of negative control group were cultured using media only containing DF12 and 10%FBS. Cells at various time points of induction were collected and expression of markers labeling retinal progenitor cells (RPCs) , photoreceptor cells (PRCs) and retinal ganglion cells (RGCs) were revealed by RT-PCR , real-time PCR and Western-blot.
Results :
The hADSCs-derived differentiated cells in the induction groups showed bright spots around nucleus, while cells in the NC group showed fibroblast-like morphology. The markers for RPCs (PAX6), PRCs (CRX, NRL and RHO), RGCs (ATOH7 and THY1), and neural precursor (NES and TUBB3) were significantly up-regulated after induction compared to negative control group.
Conclusions :
hADSCs possess the potential to differentiate toward retinal lineage by inhibiting BMP and Wnt signaling.
This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.