PMID- 15246835
OWN - NLM
STAT- MEDLINE
DCOM- 20040810
LR  - 20131121
IS  - 0014-4886 (Print)
IS  - 0014-4886 (Linking)
VI  - 188
IP  - 2
DP  - 2004 Aug
TI  - Serotonin neurons derived from rhesus monkey embryonic stem cells: similarities 
      to CNS serotonin neurons.
PG  - 351-64
AB  - We sought an in vitro primate model for serotonin neurons. Rhesus monkey 
      embryonic stem (ES) cell colonies were isolated and differentiated into embryoid 
      bodies (EBs), then transferred to serum-free medium with 1% 
      insulin-transferrin-selenium for 7 days to induce neural precursor cell (NPC) 
      formation. NPCs were cultured in medium with 1% N-2 neural supplement and human 
      fibroblast growth factor 2 (FGF2, 10 ng/ml) for 7 days to stimulate cell 
      proliferation. Lastly, NPCs were dispersed into single cells and cultured without 
      FGF2 for another 7 days to obtain terminal differentiation. Terminal cells were 
      characterized for neuronal and serotonergic markers. Over 95% of the NPCs were 
      immunopositive for nestin and Musashi1. Terminally differentiated cells appeared 
      in both small and large morphologies. Most (>95%) of the mature cells (both small 
      and large) were immunopositive for neuron-specific nuclear protein (NeuN), 
      synaptophysin, microtubule-associated protein (MAP2C), Tau-1, neurofilament 160 
      (NF-160), beta-tubulin (TujIII), tryptophan hydroxylase (TPH), serotonin, the 
      serotonin reuptake transporter (SERT), estrogen receptor-beta (ERbeta), and 
      progestin receptor (PR), but not estrogen receptor-alpha (ERalpha). Less than 
      2-3% of cells were positive for tyrosine hydroxylase (TH). Reverse transcriptase 
      polymerase chain reaction (RT-PCR) detected mRNA transcripts for TPH-1, TPH-2, 
      SERT, 5-HT1A-autoreceptor, ERbeta, and PR in the differentiated population. A low 
      level of expression of ERalpha mRNA was also detected. Quantitative RT-PCR 
      indicated that the relative abundance of TPH-2 mRNA was greater than TPH-1 mRNA. 
      Serotonin as measured by ELISA increased 3-fold in the mature stage compared to 
      the selection and expansion stages. In summary, a remarkably high percentage of 
      cells derived from monkey ES cells exhibited neuronal plus serotonergic markers 
      as well as nuclear steroid receptors similar to primate CNS serotonin neurons, 
      suggesting that these cells may serve as a useful primate model for serotonergic 
      neurons.
FAU - Salli, Ugur
AU  - Salli U
AD  - Division of Reproductive Sciences, Oregon National Primate Research Center, 
      Beaverton 97006, USA.
FAU - Reddy, Arubala P
AU  - Reddy AP
FAU - Salli, Nurgul
AU  - Salli N
FAU - Lu, Nick Z
AU  - Lu NZ
FAU - Kuo, Hung-Chih
AU  - Kuo HC
FAU - Pau, Francis K-Y
AU  - Pau FK
FAU - Wolf, Don P
AU  - Wolf DP
FAU - Bethea, Cynthia L
AU  - Bethea CL
LA  - eng
GR  - HD 18185/HD/NICHD NIH HHS/United States
GR  - MH62677/MH/NIMH NIH HHS/United States
GR  - NDS40601/DS/DS NIH HHS/United States
GR  - RR000163/RR/NCRR NIH HHS/United States
GR  - RR15199/RR/NCRR NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Exp Neurol
JT  - Experimental neurology
JID - 0370712
RN  - 0 (Antigens, Differentiation)
RN  - 0 (RNA, Messenger)
RN  - 0 (Receptors, Steroid)
RN  - 333DO1RDJY (Serotonin)
SB  - IM
MH  - Animals
MH  - Antigens, Differentiation
MH  - Cell Culture Techniques/methods
MH  - Cell Differentiation/physiology
MH  - Cell Line
MH  - Cell Lineage
MH  - Central Nervous System/*cytology
MH  - Extracellular Matrix/chemistry/metabolism
MH  - Fluorescent Antibody Technique
MH  - Gene Expression Profiling
MH  - Macaca mulatta
MH  - Neurons/*cytology/*metabolism
MH  - RNA, Messenger/biosynthesis
MH  - Receptors, Steroid/metabolism
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - Serotonin/*metabolism
MH  - Stem Cells/*cytology
EDAT- 2004/07/13 05:00
MHDA- 2004/08/11 05:00
CRDT- 2004/07/13 05:00
PHST- 2003/11/19 00:00 [received]
PHST- 2004/04/01 00:00 [revised]
PHST- 2004/04/15 00:00 [accepted]
PHST- 2004/07/13 05:00 [pubmed]
PHST- 2004/08/11 05:00 [medline]
PHST- 2004/07/13 05:00 [entrez]
AID - S0014488604001645 [pii]
AID - 10.1016/j.expneurol.2004.04.015 [doi]
PST - ppublish
SO  - Exp Neurol. 2004 Aug;188(2):351-64. doi: 10.1016/j.expneurol.2004.04.015.