Neuronal phenotypes in mouse dorsal root ganglion cell cultures: enrichment of substance P and calbindin D-28k expressing neurons in a defined medium.
The primary sensory neurons in mouse dorsal root ganglia consist of diversified subpopulations which express distinct phenotypic characteristics such as substance P or calbindin D-28k. To determine whether neuronal phenotypes are altered or not in in vitro cultures carried out in a defined synthetic medium, dissociated dorsal root ganglion cells from newborn mice were grown in the alpha-modified minimum essential medium either supplemented with 10% fetal calf serum or serum-free. About 80% of the neurons survived after 5 days of culture in both media, but only 35% or 65% were rescued after 12 days in serum-free or fetal calf serum supplemented medium, respectively. The neuronal subpopulations expressing substance P or calbindin D-28k displayed similar morphological properties in both media and a higher resistance to culture conditions than the whole neuronal cell population, especially in serum-free medium. It is therefore concluded that a defined synthetic medium offers reproducible conditions to culture dorsal root ganglion cells for at least 5 days, stimulates the expression of substance P and enriches preferentially neuronal phenotypes expressing substance P or calbindin D-28k, for a longer period of culture.
Ninomiya
T
T
Institut d'Histologie et d'Embryologie, Faculté de Médecine, Université de Lausanne, Switzerland.
Barakat-Walter
I
I
Droz
B
B
D000818:Animals; D001790:Blood Physiological Phenomena; D064026:Calbindins; D002478:Cells, Cultured; D003470:Culture Media; D016895:Culture Media, Serum-Free; D005727:Ganglia, Spinal / Q000166:cytology*; D051379:Mice; D009419:Nerve Tissue Proteins / Q000096:biosynthesis*; D009474:Neurons / Q000378:metabolism; D010641:Phenotype; D015203:Reproducibility of Results; D064030:S100 Calcium Binding Protein G / Q000096:biosynthesis*; D013373:Substance P / Q000096:biosynthesis*
D064026:Calbindins; D003470:Culture Media; D016895:Culture Media, Serum-Free; D009419:Nerve Tissue Proteins; D064030:S100 Calcium Binding Protein G; D013373:Substance P