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

J Neurosci Methods. 2017 Apr 15;282:52-60. doi: 10.1016/j.jneumeth.2017.03.005. Epub 2017 Mar 8.


Background: The 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 METHOD: hWJ-MSCs were incubated with Fast-N-Spheres medium for 24 and 72h. The new formed NSs were in turn incubated with forskolin in neurogenic NeuroForsk medium for 1-7days.

Results: hWJ-MSCs cultured with Fast-N-Spheres medium trans-differentiated into NSs in just 24h compared to 72h 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 4days 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 3-7days 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.

Conclusions: The 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.

Keywords: Forskolin; Human Wharton’s jelly; Mesenchymal stem cells; NESTIN; Neurospheres; Trans-differentiation.

MeSH terms

  • Adipogenesis / physiology
  • Blotting, Western
  • Cell Culture Techniques / methods*
  • Cell Transdifferentiation / physiology*
  • Colforsin
  • Culture Media
  • Fluorescent Antibody Technique
  • Humans
  • Karyotyping
  • Mesenchymal Stem Cells / cytology*
  • Mesenchymal Stem Cells / physiology*
  • Neurogenesis / physiology
  • Neurons / cytology*
  • Neurons / physiology*
  • Osteogenesis / physiology
  • Time Factors
  • Wharton Jelly / cytology


  • Culture Media
  • Colforsin