PMID- 25187991
OWN - NLM
STAT- MEDLINE
DCOM- 20150515
LR  - 20220716
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 9
IP  - 9
DP  - 2014
TI  - In vitro modeling of the neurovascular environment by coculturing adult human 
      brain endothelial cells with human neural stem cells.
PG  - e106346
LID - 10.1371/journal.pone.0106346 [doi]
LID - e106346
AB  - Brain and vascular cells form a functionally integrated signalling network that 
      is known as the neurovascular unit (NVU). The signalling (autocrine, paracrine 
      and juxtacrine) between different elements of this unit, especially in humans, is 
      difficult to disentangle in vivo. Developing representative in vitro models is 
      therefore essential to better understand the cellular interactions that govern 
      the neurovascular environment. We here describe a novel approach to assay these 
      cellular interactions by combining a human adult cerebral microvascular 
      endothelial cell line (hCMEC/D3) with a fetal ganglionic eminence-derived neural 
      stem cell (hNSC) line. These cell lines provide abundant homogeneous populations 
      of cells to produce a consistently reproducible in vitro model of endothelial 
      morphogenesis and the ensuing NVU. Vasculature-like structures (VLS) interspersed 
      with patches of differentiating neural cells only occurred when hNSCs were seeded 
      onto a differentiated endothelium. These VLS emerged within 3 days of coculture 
      and by day 6 were stabilizing. After 7 days of coculture, neuronal 
      differentiation of hNSCs was increased 3-fold, but had no significant effect on 
      astrocyte or oligodendrocyte differentiation. ZO1, a marker of adherens and tight 
      junctions, was highly expressed in both undifferentiated and differentiated 
      endothelial cells, but the adherens junction markers CD31 and VE-cadherin were 
      significantly reduced in coculture by approximately 20%. A basement membrane, 
      consisting of laminin, vitronectin, and collagen I and IV, separated the VLS from 
      neural patches. This simple assay can assist in elucidating the cellular and 
      molecular signaling involved in the formation of VLS, as well as the enhancement 
      of neuronal differentiation through endothelial signaling.
FAU - Chou, Chung-Hsing
AU  - Chou CH
AD  - University of Pittsburgh, McGowan Institute for Regenerative Medicine, 
      Pittsburgh, Pennsylvania, United States of America; Kings College London, 
      Institute of Psychiatry, Department of Neuroscience, London, United Kingdom; 
      Tri-service General Hospital, Department of Neurology, National Defense Medical 
      Centre, Taipei, Taiwan.
FAU - Sinden, John D
AU  - Sinden JD
AD  - ReNeuron Ltd, Guildford, Surrey, United Kingdom.
FAU - Couraud, Pierre-Olivier
AU  - Couraud PO
AD  - INSERM U1016, Institut Cochin, Paris, France; CNRS, UMR8104, Paris, France; 
      Universite Paris Descartes, Sorbonne Paris Cite, Paris, France.
FAU - Modo, Michel
AU  - Modo M
AD  - University of Pittsburgh, McGowan Institute for Regenerative Medicine, 
      Pittsburgh, Pennsylvania, United States of America; University of Pittsburgh, 
      Department of Radiology, Pittsburgh, Pennsylvania, United States of America; 
      University of Pittsburgh, Department of Bioengineering, Pittsburgh, Pennsylvania, 
      United States of America.
LA  - eng
GR  - R01 NS082226/NS/NINDS NIH HHS/United States
GR  - R01NS082226/NS/NINDS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20140904
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
SB  - IM
EIN - PLoS One. 2015;10(1):e0117650. PMID: 25635816
MH  - Astrocytes/cytology
MH  - Brain/*cytology
MH  - Cell Differentiation
MH  - Cell Line
MH  - Cells, Cultured
MH  - Coculture Techniques
MH  - Endothelial Cells/*cytology
MH  - Humans
MH  - Neural Stem Cells/*cytology
PMC - PMC4154686
COIS- Competing Interests: Dr. John Sinden is an employee of ReNeuron Ltd. The cell 
      lines described here are subject to patents, notably the D3 cell line is covered 
      under patent WO200605679A1 and the STROC05 cell line is covered under patent 
      US7666672. These cells are available through Material Transfer Agreement. This 
      does not alter the authors' adherence to all the PLOS ONE policies on sharing 
      data and materials.
EDAT- 2014/09/05 06:00
MHDA- 2015/05/16 06:00
PMCR- 2014/09/04
CRDT- 2014/09/05 06:00
PHST- 2014/02/13 00:00 [received]
PHST- 2014/08/01 00:00 [accepted]
PHST- 2014/09/05 06:00 [entrez]
PHST- 2014/09/05 06:00 [pubmed]
PHST- 2015/05/16 06:00 [medline]
PHST- 2014/09/04 00:00 [pmc-release]
AID - PONE-D-14-06494 [pii]
AID - 10.1371/journal.pone.0106346 [doi]
PST - epublish
SO  - PLoS One. 2014 Sep 4;9(9):e106346. doi: 10.1371/journal.pone.0106346. eCollection 
      2014.