Comparison of Rat Primary Midbrain Neurons Cultured in DMEM/F12 and Neurobasal Mediums
Introduction: 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 neu...
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Iran University of Medical Sciences
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doaj-229b4d763e81490b859df058ea0e8be92021-05-26T10:35:49ZengIran University of Medical SciencesBasic and Clinical Neuroscience2008-126X2228-74422021-03-01122205212Comparison of Rat Primary Midbrain Neurons Cultured in DMEM/F12 and Neurobasal MediumsNeda Valian0Mansooreh Heravi1Abolhassan Ahmadiani2Leila Dargahi3 Neuroscience Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran. Neuroscience Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran. Neuroscience Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran. Neuroscience Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran. Introduction: 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 embryos cultured in DMEM/F12 and neurobasal mediums. Methods: Primary mesencephalon cells were prepared from the E14.5 rat embryo. Then they were seeded in two different mediums ( Dulbeccochr('39')s Modified Eagle Medium/Nutrient Mixture F-12 [DMEM/F12] and neurobasal). On day 3 and day 5, half of the medium was replaced with a fresh medium. On day 7, β3-tubulin-, GFAP (Glial fibrillary acidic protein)- and Tyrosine Hydroxylase TH-positive cells were characterized as neurons, astrocytes, and dopaminergic neurons, respectively, using immunohistochemistry. Furthermore, the morphology of the cells in both mediums was observed under light microscopy on days 1, 3, and 5. Results: The cells cultured in both mediums were similar under light microscopy regarding the cell number, but in a neurobasal medium, the cells have aggregated and formed clustering structures. 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: This study’s findings demonstrated that primary midbrain cells from the E14.5 rat embryo could grow in both DMEM/F12 and neurobasal mediums. Therefore, considering the high price of a neurobasal medium, it can be replaced with DMEM/F12 for culturing primary dopaminergic neurons.http://bcn.iums.ac.ir/article-1-1379-en.htmldopaminergic neuronsrat mesencephalon cell cultureb27-supplemented neurobasaldmem/f12 medium |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Neda Valian Mansooreh Heravi Abolhassan Ahmadiani Leila Dargahi |
spellingShingle |
Neda Valian Mansooreh Heravi Abolhassan Ahmadiani Leila Dargahi Comparison of Rat Primary Midbrain Neurons Cultured in DMEM/F12 and Neurobasal Mediums Basic and Clinical Neuroscience dopaminergic neurons rat mesencephalon cell culture b27-supplemented neurobasal dmem/f12 medium |
author_facet |
Neda Valian Mansooreh Heravi Abolhassan Ahmadiani Leila Dargahi |
author_sort |
Neda Valian |
title |
Comparison of Rat Primary Midbrain Neurons Cultured in DMEM/F12 and Neurobasal Mediums |
title_short |
Comparison of Rat Primary Midbrain Neurons Cultured in DMEM/F12 and Neurobasal Mediums |
title_full |
Comparison of Rat Primary Midbrain Neurons Cultured in DMEM/F12 and Neurobasal Mediums |
title_fullStr |
Comparison of Rat Primary Midbrain Neurons Cultured in DMEM/F12 and Neurobasal Mediums |
title_full_unstemmed |
Comparison of Rat Primary Midbrain Neurons Cultured in DMEM/F12 and Neurobasal Mediums |
title_sort |
comparison of rat primary midbrain neurons cultured in dmem/f12 and neurobasal mediums |
publisher |
Iran University of Medical Sciences |
series |
Basic and Clinical Neuroscience |
issn |
2008-126X 2228-7442 |
publishDate |
2021-03-01 |
description |
Introduction: 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 embryos cultured in DMEM/F12 and neurobasal mediums.
Methods: Primary mesencephalon cells were prepared from the E14.5 rat embryo. Then they were seeded in two different mediums ( Dulbeccochr('39')s Modified Eagle Medium/Nutrient Mixture F-12 [DMEM/F12] and neurobasal). On day 3 and day 5, half of the medium was replaced with a fresh medium. On day 7, β3-tubulin-, GFAP (Glial fibrillary acidic protein)- and Tyrosine Hydroxylase TH-positive cells were characterized as neurons, astrocytes, and dopaminergic neurons, respectively, using immunohistochemistry. Furthermore, the morphology of the cells in both mediums was observed under light microscopy on days 1, 3, and 5.
Results: The cells cultured in both mediums were similar under light microscopy regarding the cell number, but in a neurobasal medium, the cells have aggregated and formed clustering structures. 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: This study’s findings demonstrated that primary midbrain cells from the E14.5 rat embryo could grow in both DMEM/F12 and neurobasal mediums. Therefore, considering the high price of a neurobasal medium, it can be replaced with DMEM/F12 for culturing primary dopaminergic neurons. |
topic |
dopaminergic neurons rat mesencephalon cell culture b27-supplemented neurobasal dmem/f12 medium |
url |
http://bcn.iums.ac.ir/article-1-1379-en.html |
work_keys_str_mv |
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