PMID- 29625493
OWN - NLM
STAT- MEDLINE
DCOM- 20180821
LR  - 20210109
IS  - 1552-5783 (Electronic)
IS  - 0146-0404 (Linking)
VI  - 59
IP  - 3
DP  - 2018 Mar 1
TI  - Characterization of a Spontaneously Immortalized Murine Muller Glial Cell Line 
      QMMuC-1.
PG  - 1666-1674
LID - 10.1167/iovs.17-23293 [doi]
AB  - PURPOSE: Muller glia are critical for the survival of retinal neurons and the 
      integrity of retinal blood vessels. Muller glial cultures are important tools for 
      investigating Muller glial pathophysiology. Here, we report a spontaneously 
      immortalized Muller glial cell line originally cultured and subsequently cloned 
      from mouse pups. The cell line, Queen's University Murine Muller glia Clone-1 
      (QMMuC-1), has been cultured for over 60 passages, has morphologic features like 
      primary Muller cell (PMC) cultures and remains stable. METHODS: QMMuC-1 and PMC 
      cells were processed for immunohistochemistry, quantitative RT-PCR, Western 
      blotting, whole cell voltage-clamping, and bioenergetic profiling. RESULTS: 
      Immunocytochemistry showed that QMMuC-1 express known Muller glial markers, 
      including glutamine synthetase, glial fibrillary acidic protein (GFAP), 
      alpha-smooth muscle actin (alpha-SMA), Aquaporin 4, Kir4.1, interleukin 33 (IL-33), 
      and sex determining region Y (SRY)-box2 (Sox2), but not Cone arrestin, Calbindin 
      1, CD68, and ionized calcium-binding adapter molecule 1 (Iba1). Compared with 
      PMC, QMMuC-1 express higher levels of chemokine (C-C motif) ligand 2 (Ccl2), 
      VEGFA, and glutamate aspartate transporter (GLAST), but lower levels of 
      interleukin 6 (IL-6), brain-derived neurotrophic factor (BDNF), insulin-like 
      growth factor 1 (IGF1), and neurotrophin 3 (NTF3). Whole-cell patch clamp 
      recordings demonstrated characteristic inward currents in response to L-glutamate 
      and L-trans-pyrrolidine-2,4-dicarboxylic acid (PDC) by QMMuC-1 cells. The 
      L-glutamate-induced current was significantly higher in QMMuC-1 cells compared 
      with PMC. Bioenergetic profiling studies revealed similar levels of glycolysis 
      and basal mitochondrial respiration between QMMuC-1 and PMC. However, 
      mitochondrial spare capacity was significantly lower in QMMuC-1 compared with 
      PMC. CONCLUSIONS: Our results suggest that the QMMuC-1 Muller glial cell line 
      retains key characteristics of PMC with its unique profiles in 
      cytokine/neurotrophic factor expression and mitochondrial respiration. QMMuC-1 
      has utility as an invaluable tool for understanding the role of Muller glia in 
      physiological and pathological conditions.
FAU - Augustine, Josy
AU  - Augustine J
AD  - Wellcome-Wolfson Institute for Experimental Medicine, Queen's University Belfast, 
      Northern Ireland, United Kingdom.
FAU - Pavlou, Sofia
AU  - Pavlou S
AD  - Wellcome-Wolfson Institute for Experimental Medicine, Queen's University Belfast, 
      Northern Ireland, United Kingdom.
FAU - O'Hare, Michael
AU  - O'Hare M
AD  - Wellcome-Wolfson Institute for Experimental Medicine, Queen's University Belfast, 
      Northern Ireland, United Kingdom.
FAU - Harkin, Kevin
AU  - Harkin K
AD  - Wellcome-Wolfson Institute for Experimental Medicine, Queen's University Belfast, 
      Northern Ireland, United Kingdom.
FAU - Stitt, Alan
AU  - Stitt A
AD  - Wellcome-Wolfson Institute for Experimental Medicine, Queen's University Belfast, 
      Northern Ireland, United Kingdom.
FAU - Curtis, Tim
AU  - Curtis T
AD  - Wellcome-Wolfson Institute for Experimental Medicine, Queen's University Belfast, 
      Northern Ireland, United Kingdom.
FAU - Xu, Heping
AU  - Xu H
AD  - Wellcome-Wolfson Institute for Experimental Medicine, Queen's University Belfast, 
      Northern Ireland, United Kingdom.
FAU - Chen, Mei
AU  - Chen M
AD  - Wellcome-Wolfson Institute for Experimental Medicine, Queen's University Belfast, 
      Northern Ireland, United Kingdom.
LA  - eng
GR  - MC_PC_15026/MRC_/Medical Research Council/United Kingdom
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Invest Ophthalmol Vis Sci
JT  - Investigative ophthalmology & visual science
JID - 7703701
RN  - 0 (Biomarkers)
RN  - 0 (Cytokines)
RN  - 0 (Nerve Growth Factors)
SB  - IM
MH  - Animals
MH  - Biomarkers/metabolism
MH  - Blotting, Western
MH  - Cell Line
MH  - Cell Membrane/physiology
MH  - Cytokines/metabolism
MH  - Ependymoglial Cells/*metabolism
MH  - Glycolysis/physiology
MH  - Immunohistochemistry
MH  - Mice
MH  - Mitochondria/metabolism
MH  - Nerve Growth Factors/metabolism
MH  - Neuroglia/*metabolism
MH  - Reverse Transcriptase Polymerase Chain Reaction
EDAT- 2018/04/07 06:00
MHDA- 2018/08/22 06:00
CRDT- 2018/04/07 06:00
PHST- 2018/04/07 06:00 [entrez]
PHST- 2018/04/07 06:00 [pubmed]
PHST- 2018/08/22 06:00 [medline]
AID - 2677080 [pii]
AID - 10.1167/iovs.17-23293 [doi]
PST - ppublish
SO  - Invest Ophthalmol Vis Sci. 2018 Mar 1;59(3):1666-1674. doi: 
      10.1167/iovs.17-23293.