PMID- 23535251
OWN - NLM
STAT- MEDLINE
DCOM- 20131212
LR  - 20130506
IS  - 1873-7544 (Electronic)
IS  - 0306-4522 (Linking)
VI  - 241
DP  - 2013 Jun 25
TI  - Development of the microglial phenotype in culture.
PG  - 280-95
LID - S0306-4522(13)00255-8 [pii]
LID - 10.1016/j.neuroscience.2013.03.033 [doi]
AB  - Selected morphological, molecular and functional aspects of various microglial 
      cell populations were characterized in cell cultures established from the 
      forebrains of E18 rat embryos. The mixed primary cortical cultures were 
      maintained for up to 28days using routine culturing techniques when the 
      microglial cells in the culture were not stimulated or immunologically 
      challenged. During culturing, expansion of the microglial cell populations was 
      observed, as evidenced by quantitative assessment of selected 
      monocyte/macrophage/microglial cell-specific markers (human leukocyte antigen 
      (HLA) DP, DQ, DR, CD11b/c and Iba1) via immunocyto- and histochemistry and 
      Western blot analysis. The Iba1 immunoreactivity in Western blots steadily 
      increased about 750-fold, and the number of Iba1-immunoreactive cells rose at 
      least 67-fold between one day in vitro (DIV1) and DIV28. Morphometric analysis on 
      binary (digital) silhouettes of the microglia revealed their evolving morphology 
      during culturing. Microglial cells were mainly ameboid in the early stages of in 
      vitro differentiation, while mixed populations of ameboid and ramified cell 
      morphologies were characteristic of older cultures as the average transformation 
      index (TI) increased from 1.96 (DIV1) to 15.17 (DIV28). Multiple 
      immunofluorescence labeling of selected biomarkers revealed different microglial 
      phenotypes during culturing. For example, while HLA DP, DQ, DR immunoreactivity 
      was present exclusively in ameboid microglia (TI<3) between DIV1 and DIV10, 
      CD11b/c- and Iba1-positive microglial cells were moderately (TI<13) and 
      progressively (TI<81) more ramified, respectively, and always present throughout 
      culturing. Regardless of the age of the cultures, proliferating microglia were 
      Ki67-positive and characterized by low TI values (TI<3). The microglial function 
      was assessed by an in vitro phagocytosis assay. Unstimulated microglia with low 
      TI values were significantly more active in phagocytosing fluorescent 
      microspheres than the ramified forms. In vitro studies on microglial population 
      dynamics combined with phenotypic characterization can be of importance when 
      different in vivo pathophysiological situations are modeled in vitro.
CI  - Copyright (c) 2013 IBRO. Published by Elsevier Ltd. All rights reserved.
FAU - Szabo, M
AU  - Szabo M
AD  - Department of Cell Biology and Molecular Medicine, University of Szeged, Szeged, 
      Hungary.
FAU - Gulya, K
AU  - Gulya K
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20130325
PL  - United States
TA  - Neuroscience
JT  - Neuroscience
JID - 7605074
SB  - IM
MH  - Animals
MH  - Blotting, Western
MH  - Cell Culture Techniques
MH  - Cells, Cultured
MH  - Embryo, Mammalian
MH  - Immunohistochemistry
MH  - Microglia/*cytology/*physiology
MH  - Phenotype
MH  - Rats
MH  - Rats, Sprague-Dawley
EDAT- 2013/03/29 06:00
MHDA- 2013/12/18 06:00
CRDT- 2013/03/29 06:00
PHST- 2012/11/23 00:00 [received]
PHST- 2013/03/12 00:00 [revised]
PHST- 2013/03/13 00:00 [accepted]
PHST- 2013/03/29 06:00 [entrez]
PHST- 2013/03/29 06:00 [pubmed]
PHST- 2013/12/18 06:00 [medline]
AID - S0306-4522(13)00255-8 [pii]
AID - 10.1016/j.neuroscience.2013.03.033 [doi]
PST - ppublish
SO  - Neuroscience. 2013 Jun 25;241:280-95. doi: 10.1016/j.neuroscience.2013.03.033. 
      Epub 2013 Mar 25.