Abstract
The short-term survival of highly purified embryonic spinal motor neurons (SMNs) in culture can be promoted by many peptide trophic factors, including brain-derived neurotrophic factor (BDNF), ciliary neurotrophic factor (CNTF), fibroblast growth factor (FGF), glial-derived neurotrophic factor (GDNF), and hepatocyte growth factor (HGF). We have asked whether these peptides are sufficient to promote the long-term survival of purified E15 SMNs. Contrary to previous reports, we find that when SMNs are cultured in serum-free medium containing a single peptide trophic factor only approximately one-third of the cells survive for 3 d in culture. When multiple factors are combined, additive effects on survival are observed transiently, but by 7 d of culture the majority of SMNs has died. Surprisingly, when cAMP levels are elevated, the majority of SMNs extend processes and survive for 1 week in culture in the absence of peptide trophic factors, even in low-density cultures. A combination of five peptide trophic factors, together with cAMP elevation, promotes the long-term survival of most of the SMNs in serum-free culture for 3 weeks. These findings provide useful culture conditions for studying the properties of SMNs and have implications for the treatment of motor neuron diseases.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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1-Methyl-3-isobutylxanthine / pharmacology
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Animals
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Bone Morphogenetic Proteins / metabolism
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Brain-Derived Neurotrophic Factor / pharmacology
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Cell Survival / physiology
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Ciliary Neurotrophic Factor
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Colforsin / pharmacology
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Cyclic AMP / metabolism*
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Estradiol / pharmacology
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Glial Cell Line-Derived Neurotrophic Factor
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Hepatocyte Growth Factor / pharmacology
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Hydrocortisone / pharmacology
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Hypoglycemic Agents / pharmacology
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Insulin / pharmacology
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Motor Neurons / cytology*
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Motor Neurons / drug effects
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Motor Neurons / metabolism*
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Nerve Degeneration / metabolism
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Nerve Growth Factors*
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Nerve Tissue Proteins / pharmacology
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Oxygen / pharmacology
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Phosphodiesterase Inhibitors / pharmacology
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Progesterone / pharmacology
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Rats
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Rats, Sprague-Dawley
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Signal Transduction / drug effects
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Signal Transduction / physiology
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Spinal Cord / cytology*
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Thyroxine / pharmacology
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Tretinoin / pharmacology
Substances
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Bone Morphogenetic Proteins
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Brain-Derived Neurotrophic Factor
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Ciliary Neurotrophic Factor
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Gdnf protein, rat
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Glial Cell Line-Derived Neurotrophic Factor
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Hypoglycemic Agents
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Insulin
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Nerve Growth Factors
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Nerve Tissue Proteins
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Phosphodiesterase Inhibitors
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Colforsin
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Progesterone
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Estradiol
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Tretinoin
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Hepatocyte Growth Factor
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Cyclic AMP
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Thyroxine
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Oxygen
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1-Methyl-3-isobutylxanthine
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Hydrocortisone