title: Astrocyte Cultures Mimicking Brain Astrocytes in Gene Expression, Signaling, Metabolism and K+Uptake and Showing Astrocytic Gene Expression Overlooked by Immunohistochemistry and In Situ Hybridization.
abstract: Based on differences in gene expression between cultured astrocytes and freshly isolated brain astrocytes it has been claimed that cultured astrocytes poorly reflect the characteristics of their in vivo counterparts. This paper shows that this is not the case with the cultures of mouse astrocytes we have used since 1978. The culture is prepared following guidelines provided by Drs. Monique Sensenbrenner and John Booher, with the difference that dibutyryl cyclic AMP is added to the culture medium from the beginning of the third week. This addition has only minor effects on glucose and glutamate metabolism, but it is crucial for effects by elevated K+concentrations and for Ca2+homeostasis, important aspects of astrocyte function. Work by Liang Peng and her colleagues has shown identity between not only gene expression but also drug-induced gene upregulations and editings in astrocytes cultured by this method and astrocytes freshly isolated from brains of drug-treated animals. Dr. Norenberg's laboratory has demonstrated identical upregulation of the cotransporter NKCC1 in ammonia-exposed astrocytes and rats with liver failure. Similarity between cultured and freshly isolated astrocytes has also been shown in metabolism, K+uptake and several aspects of signaling. However, others have shown that the gene for the glutamate transporter GLT1 is not expressed, and rat cultures show some abnormalities in K+effects. Nevertheless, the overall reliability of the cultured cells is important because immunohistochemistry and in situ hybridization poorly demonstrate many astrocytic genes, e.g., those of nucleoside transporters, and even microarray analysis of isolated cells can be misleading.
authors.affiliation: Laboratory of Brain Metabolic Diseases, Institute of Metabolic Disease Research and Drug Development, China Medical University, No. 77, Puhe Road, Shenyang North New Area, Shenyang, People's Republic of China.
authors.forename: Leif
authors.initials: L
authors.lastname: Hertz
authors.affiliation: Henry M. Jackson Foundation, Bethesda, MD, USA.
authors.forename: Ye
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Laboratory of Brain Metabolic Diseases, Institute of Metabolic Disease Research and Drug Development, China Medical University, No. 77, Puhe Road, Shenyang North New Area, Shenyang, People's Republic of China. sd_nature@hotmail.com.
authors.forename: Dan
authors.initials: D
authors.lastname: Song
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000737:chemistry / Q000378:metabolism*; D001921:Brain / Q000166:cytology / Q000378:metabolism*; D001923:Brain Chemistry / Q000502:physiology*; D002478:Cells, Cultured; D005786:Gene Expression Regulation; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D051379:Mice; D011188:Potassium / Q000378:metabolism*; D051381:Rats; D015398:Signal Transduction / Q000502:physiology*
chemical_list: D011188:Potassium
keywords: Astrocyte; Astrocytic gene expression; Cell culture; Immunohistochemistry; In situ hybridization
references.citation: Neurochem Int. 2003 Sep-Oct;43(4-5):355-61
references.pmid: 12742079
references.citation: ASN Neuro. 2014 Jan 13;6(1):e00132
references.pmid: 24328680
references.citation: J Neurosci Res. 1989 Feb;22(2):209-15
references.pmid: 2709443
references.citation: J Neurosci. 2011 Oct 5;31(40):14264-71
references.pmid: 21976511
references.citation: J Neurosci Res. 2015 Jul;93(7):999-1008
references.pmid: 25546576
references.citation: Nature. 2007 Jan 11;445(7124):168-76
references.pmid: 17151600
references.citation: J Neurochem. 2011 May;117(3):437-48
references.pmid: 21306384
references.citation: Neurobiology. 1972;2(3):97-105
references.pmid: 4572654
references.citation: Can J Physiol Pharmacol. 1992;70 Suppl:S367-73
references.pmid: 1295687
references.citation: Ann N Y Acad Sci. 1985;456:187-97
references.pmid: 2418726
references.citation: Neurochem Res. 2012 Nov;37(11):2480-95
references.pmid: 22711334
references.citation: Glia. 2014 Apr;62(4):526-34
references.pmid: 24464850
references.citation: Proc Natl Acad Sci U S A. 1994 Oct 25;91(22):10625-9
references.pmid: 7938003
references.citation: Glia. 2005 Oct;52(1):25-35
references.pmid: 15892125
references.citation: Int Rev Neurobiol. 1975;18:141-211
references.pmid: 128532
references.citation: J Neurochem. 1975 Dec;25(6):909-11
references.pmid: 173804
references.citation: Am J Physiol Cell Physiol. 2000 Dec;279(6):C1710-21
references.pmid: 11078685
references.citation: Front Neurosci. 2014 Sep 09;8:261
references.pmid: 25249930
references.citation: Brain Res. 1978 Apr 21;145(1):202-8
references.pmid: 638779
references.citation: J Physiol. 2010 Nov 1;588(Pt 21):4089-101
references.pmid: 20819947
references.citation: Neurosci Lett. 1977 Apr;4(5):263-7
references.pmid: 19604956
references.citation: Sci Signal. 2012 Apr 03;5(218):ra26
references.pmid: 22472648
references.citation: Nat Commun. 2015 Apr 23;6:6807
references.pmid: 25904018
references.citation: Dev Neurosci. 1982 Mar-Jun;5(2-3):278-84
references.pmid: 6290179
references.citation: Can J Physiol Pharmacol. 1992;70 Suppl:S165-9
references.pmid: 1295667
references.citation: J Neurochem. 1982 Jul;39(1):70-7
references.pmid: 6283023
references.citation: Neurochem Res. 2015 Feb;40(2):241-57
references.pmid: 24929663
references.citation: Am J Physiol Cell Physiol. 2012 Feb 1;302(3):C505-17
references.pmid: 22049209
references.citation: Cell. 2011 Mar 4;144(5):810-23
references.pmid: 21376239
references.citation: Neurochem Res. 2012 Nov;37(11):2524-40
references.pmid: 22965852
references.citation: Glia. 1997 Sep;21(1):35-45
references.pmid: 9298845
references.citation: Brain Res. 1984 Feb 6;292(2):367-74
references.pmid: 6692162
references.citation: J Neurophysiol. 2002 Jan;87(1):87-102
references.pmid: 11784732
references.citation: J Neurosci Res. 1986;16(4):699-708
references.pmid: 3025461
references.citation: PLoS One. 2011;6(12):e28427
references.pmid: 22180782
references.citation: Fundam Clin Pharmacol. 2014 Apr;28(2):217-20
references.pmid: 23130601
references.citation: Neurochem Int. 2010 Nov;57(4):411-20
references.pmid: 20380860
references.citation: J Cereb Blood Flow Metab. 1992 Jan;12(1):34-42
references.pmid: 1727141
references.citation: Neurochem Res. 2015 Dec;40(12 ):2493-504
references.pmid: 26168779
references.citation: Neurochem Res. 1981 Apr;6(4):353-64
references.pmid: 6267494
references.citation: J Neurosci. 2011 Mar 23;31(12):4709-13
references.pmid: 21430169
references.citation: J Neurochem. 1973 Jul;21(1):191-8
references.pmid: 4720894
references.citation: Neurochem Res. 2015 Dec;40(12):2505-16
references.pmid: 25555706
references.citation: Brain Res. 1976 Jun 11;109(2):311-22
references.pmid: 1276917
references.citation: Front Behav Neurosci. 2015 Feb 20;9:25
references.pmid: 25750618
references.citation: Neurochem Res. 2013 Nov;38(11):2351-8
references.pmid: 24026568
references.citation: J Neurochem. 1979 Jun;32(6):1787-92
references.pmid: 221617
references.citation: Proc Natl Acad Sci U S A. 1995 May 9;92(10):4616-20
references.pmid: 7753851
references.citation: Neurochem Int. 2012 Jul;61(2):187-94
references.pmid: 22564531
references.citation: J Neurosci Res. 2007 Nov 15;85(15):3367-77
references.pmid: 17847082
references.citation: J Neurochem. 1985 Mar;44(3):723-30
references.pmid: 2983017
references.citation: Neurochem Res. 2013 Nov;38(11):2364-74
references.pmid: 24037520
references.citation: Glia. 1995 Oct;15(2):152-6
references.pmid: 8567066
references.citation: Neurochem Res. 2002 Feb;27(1-2):113-20
references.pmid: 11930908
references.citation: Brain Res. 1980 Oct 20;199(2):493-500
references.pmid: 7417799
references.citation: J Hepatol. 2011 Feb;54(2):272-8
references.pmid: 21056502
references.citation: Glia. 2014 Apr;62(4):608-22
references.pmid: 24482245
references.citation: J Cereb Blood Flow Metab. 2011 Jan;31(1):90-101
references.pmid: 20736955
references.citation: Brain Res Dev Brain Res. 1994 Mar 18;78(1):137-41
references.pmid: 8004768
references.citation: Adv Exp Med Biol. 1990;265:227-43
references.pmid: 1974106
references.citation: J Neurophysiol. 1997 Jul;78(1):461-77
references.pmid: 9242294
references.citation: J Neurochem. 1977 May;28(5):897-916
references.pmid: 864466
references.citation: J Neurosci Res. 2006 Sep;84(4):801-8
references.pmid: 16862552
references.citation: Glia. 2002 Feb;37(2):114-23
references.pmid: 11754210
references.citation: Front Integr Neurosci. 2013 Apr 03;7:20
references.pmid: 23565080
references.citation: J Cell Biol. 1980 Jun;85(3):890-902
references.pmid: 6248568
references.citation: Neurochem Int. 2012 Dec;61(8):1325-32
references.pmid: 23017600
references.citation: J Signal Transduct. 2014;2014:593934
references.pmid: 24707399
references.citation: J Physiol. 2000 Feb 1;522 Pt 3:427-42
references.pmid: 10713967
references.citation: Curr Top Med Chem. 2011;11(8):948-72
references.pmid: 21401500
references.citation: Neurochem Res. 1978 Feb;3(1):1-14
references.pmid: 683409
references.citation: Brain Res Bull. 1995;38(2):153-9
references.pmid: 7583341
references.citation: Brain Res. 1985 Aug 12;340(2):333-40
references.pmid: 2411355
references.citation: Neurol Res. 2005 Apr;27(3):280-6
references.pmid: 15845211
references.citation: J Comput Neurosci. 2008 Oct;25(2):349-65
references.pmid: 18297383
references.citation: Can J Physiol Pharmacol. 1992;70 Suppl:S138-44
references.pmid: 1295664
references.citation: Neurochem Res. 2014 Apr;39(4):661-7
references.pmid: 24500447
references.citation: J Neuropathol Exp Neurol. 1993 May;52(3):205-15
references.pmid: 8388039
references.citation: Glia. 2006 Aug 15;54(3):214-22
references.pmid: 16819764
references.citation: Am J Physiol Regul Integr Comp Physiol. 2006 Jul;291(1):R1-25
references.pmid: 16484438
references.citation: J Neurosci Methods. 2011 Jan 15;194(2):206-17
references.pmid: 20951737
references.citation: J Cereb Blood Flow Metab. 1984 Jun;4(2):301-4
references.pmid: 6725441
references.citation: ASN Neuro. 2012 Mar 22;4(2):null
references.pmid: 22300409
references.citation: Neurochem Int. 2012 Sep;61(4):566-74
references.pmid: 22306776
references.citation: Neuroscientist. 2010 Feb;16(1):79-106
references.pmid: 20236950
references.citation: Neuron. 2008 Sep 25;59(6):932-46
references.pmid: 18817732
references.citation: Adv Neurobiol. 2014;11:31-54
references.pmid: 25236723
references.citation: Neurochem Pathol. 1988 Jul-Dec;9:195-209
references.pmid: 3247069
references.citation: Neuropathology. 2011 Jun;31(3):250-64
references.pmid: 21118399
references.citation: Semin Nephrol. 1987 Sep;7(3):185-92
references.pmid: 2446366
references.citation: Neurochem Res. 2012 Nov;37(11):2299-309
references.pmid: 22367475
references.citation: Biomed Res Int. 2014;2014:873590
references.pmid: 25478577
references.citation: J Neurosci. 2007 Nov 7;27(45):12255-66
references.pmid: 17989291
references.citation: J Neurosci. 2008 Jan 2;28(1):264-78
references.pmid: 18171944
references.citation: Neurochem Res. 2012 Nov;37(11):2569-88
references.pmid: 22926576
references.citation: J Physiol. 1975 Oct;252(1):115-36
references.pmid: 1202194
references.citation: Magn Reson Imaging. 2011 Dec;29(10):1319-29
references.pmid: 21820830
references.citation: Biochem Biophys Res Commun. 2005 Sep 9;334(4):1343-50
references.pmid: 16043124
references.citation: Front Endocrinol (Lausanne). 2013 Sep 17;4:123
references.pmid: 24062726
references.citation: Adv Anat Embryol Cell Biol. 1975;52(1):3-89
references.pmid: 813499
references.citation: J Neuropathol Exp Neurol. 2004 Aug;63(8):856-71
references.pmid: 15330340
references.citation: J Biol Chem. 2008 Dec 5;283(49):33874-82
references.pmid: 18849345
references.citation: Neurochem Int. 2000 Aug-Sep;37(2-3):163-70
references.pmid: 10812201
references.citation: FEBS Lett. 1995 Jul 3;367(3):319-25
references.pmid: 7607331
references.citation: Physiol Bohemoslov. 1979;28(2):105-11
references.pmid: 223182
references.citation: J Neurochem. 2003 Apr;85(1):73-81
references.pmid: 12641728
references.citation: J Cereb Blood Flow Metab. 2004 Sep;24(9):1004-14
references.pmid: 15356421
references.citation: Neuron. 2011 Sep 8;71(5):799-811
references.pmid: 21903074
references.citation: J Cereb Blood Flow Metab. 2013 Apr;33(4):550-6
references.pmid: 23281428
references.citation: Exp Brain Res. 1970;11(2):199-212
references.pmid: 5472272
references.citation: Front Endocrinol (Lausanne). 2013 Nov 25;4:176
references.pmid: 24324456
references.citation: Brain Res. 1987 Apr 28;410(1):164-8
references.pmid: 3034384
references.citation: Metab Brain Dis. 2015 Feb;30(1):317-33
references.pmid: 24744118
references.citation: Nature. 1976 Dec 9;264(5586):560-2
references.pmid: 1004595
references.citation: Dev Neurosci. 1986;8(1):31-43
references.pmid: 3017675
references.citation: J Neurochem. 1988 Apr;50(4):1179-84
references.pmid: 3346673
references.citation: Neuron. 2009 Mar 26;61(6):820-38
references.pmid: 19323993
references.citation: Brain Res. 1978 Sep 15;153(1):55-77
references.pmid: 209876
references.citation: Br J Pharmacol. 2008 May;154(1):191-203
references.pmid: 18311185
references.citation: Brain Res. 1980 Jan 13;181(2):482-7
references.pmid: 7350977
references.citation: J Neurotrauma. 2014 Jul 15;31(14):1249-57
references.pmid: 24471369
references.citation: Glia. 2001 Dec;36(3):330-41
references.pmid: 11746770
references.citation: J Neurosci Res. 2007 Nov 15;85(15):3326-33
references.pmid: 17455305
references.citation: In Vitro. 1979 Jun;15(6):429-36
references.pmid: 478567
references.citation: Front Cell Neurosci. 2014 Nov 06;8:378
references.pmid: 25414643
references.citation: Biochim Biophys Acta. 1987 Oct 16;903(3):411-6
references.pmid: 2444257
references.citation: ASN Neuro. 2014 Jul 22;6(4):1759091414543439
references.pmid: 25298788
references.citation: J Biol Chem. 2007 Oct 5;282(40):29414-23
references.pmid: 17684014
references.citation: Neurochem Res. 2015 Dec;40(12):2402-13
references.pmid: 25894677
references.citation: J Neurosci. 1988 Jun;8(6):1922-8
references.pmid: 3385482
references.citation: J Neurophysiol. 2000 Mar;83(3):1443-51
references.pmid: 10712471
references.citation: Cereb Cortex. 2008 Dec;18(12):2789-95
references.pmid: 18372288
references.citation: J Neurosci Res. 1980;5(6):497-506
references.pmid: 6259370
references.citation: Experientia. 1976 Jan 15;32(1):40-2
references.pmid: 174936
references.citation: Neurochem Int. 1996 Mar;28(3):335-42
references.pmid: 8813252
references.citation: Neurochem Res. 2010 Sep;35(9):1434-44
references.pmid: 20577800
references.citation: Dev Neurosci. 1996;18(5-6):353-9
references.pmid: 8940606
references.citation: Glia. 1991;4(5):448-55
references.pmid: 1718862
references.citation: Neurochem Res. 2012 Nov;37(11):2613-26
references.pmid: 23079895
references.citation: J Neurochem. 2010 Nov;115(4):1007-23
references.pmid: 20831657
references.citation: Metab Brain Dis. 2015 Feb;30(1):281-98
references.pmid: 24515302
references.citation: Brain Res Dev Brain Res. 2003 Jun 12;143(1):33-46
references.pmid: 12763579
references.citation: J Neurosci Res. 2015 Jul;93(7):1019-30
references.pmid: 25352321
references.citation: J Neurosurg. 1990 Dec;73(6):889-900
references.pmid: 1977896
references.citation: J Physiol. 1976 Jan;254(1):22P-23P
references.pmid: 1249732