Fast transdifferentiation of human Wharton's jelly mesenchymal stem cells into neurospheres and nerve-like cells

- Neurospheres (NSs) are obtained by 24 h transdifferentiation.
- Fast-N-Spheres medium favors NSs NESTIN positive formation.
- NSs expressed low NANOG levels and SOX2 and OCT4 are undetected.
- Nerve-like cells (NLCs) are obtained by exposing NSs to NeuroForsk medium by day 4.
- NLCs expressed neuronal markers MAP-2, NF-L, TUC-4 and β-TUB-III.

BackgroundThe human mesenchymal stem cells derived from Wharton's jelly tissue (hWJ-MSCs) represent a tool for cell-based therapies and regenerative medicine. hWJ-MSCs form neurospheres (NSs) within 3-7 days. No data is available to establish the neuro-phenotypic markers and time of formation of nerve-like (NLCs) and glial cells from NSs derived from hWJ-MSCs.New methodhWJ-MSCs were incubated with Fast-N-Spheres medium for 24 and 72 h. The new formed NSs were in turn incubated with forskolin in neurogenic NeuroForsk medium for 1-7 days.ResultshWJ-MSCs cultured with Fast-N-Spheres medium trans-differentiated into NSs in just 24 h compared to 72 h for hWJ-MSCs cultured with classic growth factor medium. The NSs generated from the Fast-N-Spheres medium expressed reduced levels SOX2, OCT4 and NANOG, as markers of pluripotency compared to undifferentiated hWJ-MSCs. The formed NSs exposed to NeuroForsk medium differentiated into NLCs in 4 days as evidenced by high levels of protein expression of the neuronal markers, and no expression of the glial marker GFAP.Comparison with existing method(s)Currently, the formation and harvest of NSs is expensive and time consuming. Published protocols require 3-7 days to form NSs from whole human umbilical cord MSCs. We report for the first time, to our knowledge, the differentiation of NSs-derived from hWJ-MSCs into NLCs.ConclusionsThe fastest method to obtain NSs and NLCs from hWJ-MSCs takes only five days using the two-step incubation media Fast-N-Spheres and NeuroForsk.

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