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Preprints

Neural differentiation footprints of mesenchymal stem cell by fetal brain extract

Razeghian-Jahromi I, Mehrabani D, Mohammadi A, Seno MMG, Zare S, Zibaeenezhad MJ, Mowla A.
Preprint from
Research Square
7 May 2020
PPR
PPR159713
Abstract

Background:

Stem cells has brought promising hope to improve impairment in different organs especially those with limited intrinsic regeneration capacity like nervous system. With the use of mesenchymal stem cells' (MSC) capacity to differentiate toward neural cells, this study aimed to examine the potential of fetal rat brain extract (FBE) as a biological inducer to mimic natural differentiation environment. Methods Bone marrow-MSCs were derived from Rattus norvegicus . After characterization of their mesenchymal nature, three groups were considered including control (culture medium only), differentiation (culture medium supplemented by basic fibroblast growth factor and epidermal growth factor), and treatment (culture medium and FBE). Growth curves as well as expression of glial fibrillary acidic protein (GFAP, marker for astroglial cells), galactocerebridase (GaLc, marker for oligodendrocyte and schwann cells), choline acetyl transferase (ChAT, marker for cholinergic neurons), and tyrosine hydroxylase (TH, marker of dopaminergic neurons) were examined in all the three groups at day 3 and 7 post-exposure. Also, nissl staining was performed in the three groups in order to assess protein synthesis and neurotransmitter production. Results Spindle-shaped cells were seen in the control group. There were neural-like cells in the differentiation group while more developed appearance with axon- and dendrite-like processes were present in the treatment group. To examine cell viability after exposure to FBE, growth curve was delineated. Higher proliferation rate and lower population doubling time were seen in the treatment group compared to other two groups. Regarding expression of neural markers, GaLC and ChAT at day 7 was prominent in the treatment group. Nissl bodies, which are rough endoplasmic reticulum, were seen only in the treatment group. Conclusions FBE might be a safer and more efficient inducer than combination of two well-known growth factors in terms of differentiation of MSCs to functional dopaminergic neurons. Making neural cells in vitro is an attractive approach in improvement of injured nervous system.