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J. Ge, Y. Qi, B.-Q. Liu, X.-R. Sun, R.-Z. Jiang, M.-F. Chen; Differentiation of Rhesus Monkey BMSCs Into Neuron-Like Cells Induced With Sonic Hedgehog and Retinoic Acid. Invest. Ophthalmol. Vis. Sci. 2009;50(13):1294.
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© ARVO (1962-2015); The Authors (2016-present)
To investigate the efficacy of sonic hedgehog (SHH), retinoic acid (RA), and (SHH+RA) on inducing the differentiation of rhesus monkey BMSCs into neurons.
Rhesus monkey BMSCs were isolated, purified and expanded. Passages 3-5 were used in this study. Specific markers were detected with flow cytometry. Neuronal inducing medium contains 10 ng/ml bFGF, 2% B27, 5µM Forskolin, 125 µM IBMX, and 50 ng/ml BDNF dissolved in DMEM, and supplemented with one of the following reagents: a. 250 ng/ml SHH, b. 0.5µM RA, c. SHH+RA, d.vehicle (control). The inducing mediums were incubated with the BMSCs for additional 3 days in each group. The expressions of nestin, NSE, synaptophysin and GFAP were tested at different time points using RT-PCR, immunofluorescence, and Western blot.
Before induction, markers CD29, CD44, and CD105 were positive in BMSCs detected by flow cytometry, while CD34, CD45, and CD11b negative. From day 1, the morphology of induced cells changed dramatically and evenly in SHH, RA, and SHH+RA groups, but not in control. After 3 days of induction, nestin, SOX2, and NSE expression showed positive by immunofluorescence and western blot assays. The difference between each of the three groups and vehicle control was significant, but there was no statistical difference among groups a, b and c.
SHH and/or RA could induce rhesus monkey BMSCs to differentiate into neuron-like cells. Further study on the differentiation mechanism and function restoration might facilitate the regeneration of retina neurons.
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