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1- Neuroscience Research Center, Shahid Beheshti University of Medical Sciences
2- Neurobiology Research Center, Shahid Beheshti University of Medical Sciences
Background: Midbrain dopaminergic neurons are involved in various brain functions including motor behavior, reinforcement, motivation, learning and cognition. Primary dopaminergic neurons and also several lines of these cells are extensively used in cell culture studies. Primary dopaminergic neurons prepared from rodents have been cultured in both DMEM/F12 and neurobasal mediums in several studies. However there is no document reporting the comparison of these two mediums. So in this study we evaluated the neurons and astroglial cells in primary midbrain neurons from rat embryo cultured in DMEM/F12 and neurobasal mediums.
Methods: Primary mesencephalon cells were prepared from E14.5 rat embryo and were seeded in two different mediums (DMEM/F12 and neurobasal). On day 3 and day 5 the half of medium were replaced with fresh medium. On day 7, β3-tubulin-, GFAP- and TH-positive cells were characterized as neurons, astrocytes and dopaminergic neurons respectively, using immunohistochemistry. Furthermore, the morphology of the cells in both mediums observed under light microscopy on day 1, 3 and 5.
Results: The cells cultured in both mediums were similar under light microscopy regarding the cell number, but in neurobasal medium the cells has aggregated and formed clustering structure. Although GFAP-immunoreactive cells were lower in neurobasal compared to DMEM/F12, the number of β3-tubulin- and TH-positive cells in both cultures was the same.
Conclusion: Finding of this study demonstrated that primary midbrain cells from E14.5 rat embryo can grow in both DMEM/F12 and neurobasal mediums. Therefore, considering the costly-price of neurobasal medium, it can be replaced with DMEM/F12 for culturing primary dopaminergic neurons.
Type of Study: Original | Subject: Cellular and molecular Neuroscience
Received: 2018/12/10 | Accepted: 2019/05/13 | Published: 2018/03/15

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