RRC ID |
43724
|
Author |
Shimada T, Takai Y, Shinohara K, Yamasaki A, Tominaga-Yoshino K, Ogura A, Toi A, Asano K, Shintani N, Hayata-Takano A, Baba A, Hashimoto H.
|
Title |
A simplified method to generate serotonergic neurons from mouse embryonic stem and induced pluripotent stem cells.
|
Journal |
J Neurochem
|
Abstract |
We have developed a new simple method to induce serotonergic neurons from embryonic stem (ES) and induced pluripotent stem cells. When ES or induced pluripotent stem cells were cultured on a thick gel layer of Matrigel, most colonies extended TuJ1-positive neurites. We found that noggin, a known antagonist of bone morphogenic protein, induces ES cells to express genes involved in serotonergic differentiation, such as Nkx2.2, Pet-1, Sonic hedgehog, tryptophan hydroxylase 2, and serotonin transporter, as well as increases high potassium-induced release of serotonin. To concentrate serotonergic neurons, ES cells carrying Pet-1-enhancer-driven enhanced green fluorescent protein were differentiated and sorted into about 80% pure cultures of serotonergic neurons. Whole cell voltage-clamp recordings showed a voltage-dependent current in dissociated neurons. This simplified method provides an alternative option for serotonergic differentiation of pluripotent stem cells and will likely contribute a deeper understanding regarding the nature of serotonergic neurons and open new therapeutic perspectives for the treatment of psychiatric disorders.
|
Volume |
122(1)
|
Pages |
81-93
|
Published |
2012-7-1
|
DOI |
10.1111/j.1471-4159.2012.07724.x
|
PMID |
22404309
|
MeSH |
Animals
Bone Morphogenetic Protein 4 / pharmacology
Carrier Proteins / metabolism
Carrier Proteins / pharmacology
Cell Differentiation / physiology
Cell Line
Collagen / metabolism
Dose-Response Relationship, Drug
Drug Combinations
Electric Stimulation
Embryo, Mammalian
Embryonic Stem Cells / physiology*
Flow Cytometry
Green Fluorescent Proteins / genetics
Hippocampus / cytology
Homeobox Protein Nkx-2.2
Induced Pluripotent Stem Cells / physiology*
Laminin / metabolism
Mice
Organ Culture Techniques
Proteins / genetics
Proteins / metabolism
Proteoglycans / metabolism
RNA, Messenger / metabolism
RNA, Untranslated
Serotonergic Neurons / physiology*
Serotonin / metabolism
Transcription Factors / genetics
Transduction, Genetic
Tubulin / metabolism
Tyrosine 3-Monooxygenase / metabolism
|
IF |
4.066
|
Times Cited |
21
|
WOS Category
|
BIOCHEMISTRY & MOLECULAR BIOLOGY
NEUROSCIENCES
|
Resource |
Human and Animal Cells |
MC3T3-G2/PA6(RCB1127) |