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Background: Human bone marrow mesenchymal stem cells (hMSCs)
can differentiate into several types of mesenchymal cells, including
osteocytes, chondrocytes, and adipocytes, but can also differentiate into
non-mesenchymal cells, such as neural cells, under appropriate experimental
conditions. Until now, many protocols for inducing neuro-differentiation in MSCs
in vitro have been reported. In this study, we induced differentiation into
neural phenotype in the hMSCs population by new
protocol. In this treatment, hMSCs could express
neural markers more than other reports, associating with remarkable
morphological modifications.
Methods: The Bone marrow specimens were aspirated from the iliac crest of normal men. hMSCs
were isolated and cultured in DMEM containing 15%
FBS. Between 4-8
passages conversion of hMSCs into
neurosphere-like structures and induction this cells to nerve precursors in the
low-attachment plastic bacterial dishes with bFGF,
EGF & RA
were initiated. After seven days terminal neural differentiation was initiated
by plating the cells on poly-L-ornithin and Laminin
coated dishes. Cells were
differentiated for 7-14 days. We used
flowcytometry and immunocytochemistry analysis for assessment of specific
neural stem cell markers in induced cells.
Results: Flowcytometery analysis showed that after induction, 90±2.52
percent of the cells will express neuronal marker Nestin and about 41±1
percent of the cells will express Tuj-1
and about 67±1.05 percent of the cells will
express GFAP. Immunocytochemistry
and morphologically modifications revealed the same results.
Conclusion: Results
showed that hMSCs treatment with bFGF, EGF
& RA the number of Tuj1
neurons. These data confirmed that hMSCs
can exhibit neuronal differentiation potential in vitro, depending on the
protocols of inducement.
