In Vitro Effects of Wistar Rat Prenatal and Postnatal Cerebrospinal Fluid on Neural Differentiation and P roliferation of Mesenchymal Stromal Cells Derived from Bone Marrow.
Objective: Cerebrospinal fluid (CSF) plays an important role in cortical development during the fetal stages. Embryonic
CSF (E-CSF) consists of numerous neurotrophic and growth factors that regulate neurogenesis, differentiation, and
proliferation. Mesenchymal stem cells (MSCs) are multi-potential stem cells that can differentiate into mesenchymal
and non-mesenchymal cells, including neural cells. This study evaluates the prenatal and postnatal effects of CSF on
proliferation and neural differentiation of bone marrow MSCs (BM-MSCs) at gestational ages E19, E20, and the first
day after birth (P1).
Materials and Methods: In this experimental study, we confirmed the mesenchymal nature of BM-MSCs according to
their adherence properties and surface markers (CD44, CD73 and CD45). The multi-potential characteristics of BMMSCs
were verified by assessments of the osteogenic and adipogenic potentials of these cells. Under appropriate in
vitro conditions, the BM-MSCs cultures were incubated with and without additional pre- and postnatal CSF. The MTT
assay was used to quantify cellular proliferation and viability. Immunocytochemistry was used to study the expression
of MAP-2 and β-III tubulin in the BM-MSCs. We used ImageJ software to measure the length of the neurites in the
cultured cells.
Results: BM-MSCs differentiated into neuronal cell types when exposed to basic fibroblast growth factor (b-FGF).
Viability and proliferation of the BM-MSCs conditioned with E19, E20, and P1 CSF increased compared to the control
group. We observed significantly elevated neural differentiation of the BM-MSCS cultured in the CSF-supplemented
medium from E19 compared to cultures conditioned with E20 and P1 CSF group.
Conclusion: The results have confirmed that E19, E20, and P1 CSF could induce proliferation and differentiation of
BM-MSCs though they are age dependent factors. The presented data support a significant, conductive role of CSF
components in neuronal survival, proliferation, and differentiation
