PMID- 24084642
OWN - NLM
STAT- MEDLINE
DCOM- 20140410
LR  - 20220517
IS  - 1940-087X (Electronic)
IS  - 1940-087X (Linking)
IP  - 79
DP  - 2013 Sep 16
TI  - Laser capture microdissection of neurons from differentiated human 
      neuroprogenitor cells in culture.
PG  - e50487
LID - 10.3791/50487 [doi]
LID - 50487
AB  - Neuroprogenitor cells (NPCs) isolated from the human fetal brain were expanded 
      under proliferative conditions in the presence of epidermal growth factor (EGF) 
      and fibroblast growth factor (FGF) to provide an abundant supply of cells. NPCs 
      were differentiated in the presence of a new combination of nerve growth factor 
      (NGF), brain-derived neurotrophic factor (BDNF), dibutyryl cAMP (DBC) and 
      retinoic acid on dishes coated with poly-L-lysine and mouse laminin to obtain 
      neuron-rich cultures. NPCs were also differentiated in the absence of 
      neurotrophins, DBC and retinoic acid and in the presence of ciliary neurotrophic 
      factor (CNTF) to yield astrocyte-rich cultures. Differentiated NPCs were 
      characterized by immunofluorescence staining for a panel of neuronal markers 
      including NeuN, synapsin, acetylcholinesterase, synaptophysin and GAP43. Glial 
      fibrillary acidic protein (GFAP) and STAT3, astrocyte markers, were detected in 
      10-15% of differentiated NPCs. To facilitate cell-type specific molecular 
      characterization, laser capture microdissection was performed to isolate neurons 
      cultured on polyethylene naphthalate (PEN) membrane slides. The methods described 
      in this study provide valuable tools to advance our understanding of the 
      molecular mechanism of neurodegeneration.
FAU - Bouchard, Ron
AU  - Bouchard R
AD  - Section of Endocrinology, Denver VA Medical Center.
FAU - Chong, Thomas
AU  - Chong T
FAU - Pugazhenthi, Subbiah
AU  - Pugazhenthi S
LA  - eng
GR  - I01 BX001837/BX/BLRD VA/United States
GR  - I01 BX004480/BX/BLRD VA/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PT  - Video-Audio Media
DEP - 20130916
PL  - United States
TA  - J Vis Exp
JT  - Journal of visualized experiments : JoVE
JID - 101313252
RN  - EC 3.1.- (Endonucleases)
RN  - EC 3.1.30.- (CEL I nuclease)
SB  - IM
MH  - Brain/cytology/embryology
MH  - Cell Differentiation/physiology
MH  - Endonucleases/physiology
MH  - Fetus/cytology
MH  - Fluorescent Antibody Technique/methods
MH  - Humans
MH  - Laser Capture Microdissection/*methods
MH  - Neural Stem Cells/*cytology
MH  - Neurons/*cytology
MH  - Staining and Labeling/methods
PMC - PMC3885972
EDAT- 2013/10/03 06:00
MHDA- 2014/04/11 06:00
PMCR- 2015/09/16
CRDT- 2013/10/03 06:00
PHST- 2013/10/03 06:00 [entrez]
PHST- 2013/10/03 06:00 [pubmed]
PHST- 2014/04/11 06:00 [medline]
PHST- 2015/09/16 00:00 [pmc-release]
AID - 50487 [pii]
AID - 10.3791/50487 [doi]
PST - epublish
SO  - J Vis Exp. 2013 Sep 16;(79):e50487. doi: 10.3791/50487.