Preliminary Study On In Vitro Culture Of Ventral Mesencephalic Precursors From Inbred Rat Embryos And The Relative Neurotrophic Effect Of Bone Marrow Stromal Cells

Objective: To establish a foundation for studying the molecular differentiation mechanism of dopaminergic neurons (DNs) from ventral mesencephalic precursors (VMPs), by means of culturing VMPs from inbred rat embryos in vitro.Methods: VMPs harvested from ventral mesencephalons of E11 inbred rat embryos were cultured in the medium supplemented with bFGF for seven days, which subsequently differentiated into DNs in special free-bFGF medium containing AA-2P during the second seven days. The growth curve of these cells was made on the basis of close observation on VMPs with an inversion microscope. The yields of differentiated cells were identified by immunocytochemical staining and flow cytometry.Results: The number of VMPs cultured for fourteen days multiplied about 49.76±6.34 folds. Immunocytochemical staining showed that about (24.85± 12.85)% of the total cell population differentiated into Tyrosine Hydroxylase (TH)-positive dopaminergic neurons which comprised about (71.33±20.42)% of all the neurons. Flow cytometry also demonstrated similar results.Conclusion: VMPs from inbred rat embryos can differentiate into abundance of DNs through culture in vitro, which therefore can be used as an ideal cell model for studying the molecular mechanism underlying the differentiation from VMPs into DNs.