Amyloid precursor protein and proinflammatory changes are regulated in brain and adipose tissue in a murine model of high fat diet-induced obesity

PLoS ONE

Volumn 7, Issue 1, 2012, Pages

  Puig, Kendra L ^a   Floden, Angela M ^a   Adhikari, Ramchandra ^a   Golovko, Mikhail Y ^a   Combs, Colin K ^a  

^a UNIVERSITY OF NORTH DAKOTA SCHOOL OF MEDICINE AND HEALTH SCIENCES   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

AMYLOID PRECURSOR PROTEIN; MEMBRANE PROTEIN;

ABDOMINAL FAT; ADIPOCYTE; ADIPOSE TISSUE; ANIMAL CELL CULTURE; ANIMAL EXPERIMENT; ANIMAL MODEL; ANIMAL TISSUE; ARTICLE; BRAIN; CELL ACTIVATION; CONTROLLED STUDY; HIGH FAT DIET INDUCED OBESITY; HIPPOCAMPUS; IMMUNOCOMPETENT CELL; IN VIVO STUDY; INTRAABDOMINAL FAT; MACROPHAGE ACTIVATION; MICROGLIA; MOUSE; NONHUMAN; OBESITY; PHENOTYPE; PROTEIN EXPRESSION; RISK FACTOR; SUBCUTANEOUS FAT; ANIMAL; C57BL MOUSE; CHEMICALLY INDUCED DISORDER; DRUG EFFECT; INFLAMMATION; LIPID DIET; MALE; METABOLISM;

ANIMALIA; MURINAE; MUS;

ADIPOSE TISSUE; AMYLOID BETA-PROTEIN PRECURSOR; ANIMALS; BRAIN; DIET, HIGH-FAT; INFLAMMATION; MALE; MICE; MICE, INBRED C57BL; OBESITY;

EID: 84856021238     PISSN: None     EISSN: 19326203     Source Type: Journal    
DOI: 10.1371/journal.pone.0030378     Document Type: Article

References (102)

1. Kivipelto M, Ngandu T, Fratiglioni L, Viitanen M, Kareholt I, et al. (2005) Obesity and vascular risk factors at midlife and the risk of dementia and Alzheimer disease. Archives of Neurology 62: 1556-1560.
2. Beydoun MA, Beydoun HA, Wang Y, (2008) Obesity and central obesity as risk factors for incident dementia and its subtypes: a systematic review and meta-analysis. Obesity Reviews 9: 204-218.
3. Fitzpatrick AL, Kuller LH, Lopez OL, Diehr P, O'Meara ES, et al. (2009) Midlife and late-life obesity and the risk of dementia: cardiovascular health study. Archives of Neurology 66: 336-342.
4. Whitmer RA, (2007) The epidemiology of adiposity and dementia. Current Alzheimer research 4: 117-122.
5. Whitmer RA, Gunderson EP, Quesenberry CP Jr, Zhou J, Yaffe K, (2007) Body mass index in midlife and risk of Alzheimer disease and vascular dementia. Current Alzheimer research 4: 103-109.
6. Profenno LA, Faraone SV, (2008) Diabetes and overweight associate with non-APOE4 genotype in an Alzheimer's disease population. Am J Med Genet B Neuropsychiatr Genet 147B: 822-829.
7. Profenno LA, Porsteinsson AP, Faraone SV, (2010) Meta-analysis of Alzheimer's disease risk with obesity, diabetes, and related disorders. Biol Psychiatry 67: 505-512.
8. Laitinen MH, Ngandu T, Rovio S, Helkala EL, Uusitalo U, et al. (2006) Fat intake at midlife and risk of dementia and Alzheimer's disease: a population-based study. Dement Geriatr Cogn Disord 22: 99-107.
9. Eskelinen MH, Ngandu T, Helkala EL, Tuomilehto J, Nissinen A, et al. (2008) Fat intake at midlife and cognitive impairment later in life: a population-based CAIDE study. Int J Geriatr Psychiatry 23: 741-747.
10. Razay G, Vreugdenhil A, Wilcock G, (2006) Obesity, abdominal obesity and Alzheimer disease. Dementia and geriatric cognitive disorders 22: 173-176.
11. Razay G, Vreugdenhil A, Wilcock G, (2007) The metabolic syndrome and Alzheimer disease. Arch Neurol 64: 93-96.
12. Raffaitin C, Gin H, Empana JP, Helmer C, Berr C, et al. (2009) Metabolic syndrome and risk for incident Alzheimer's disease or vascular dementia: the Three-City Study. Diabetes Care 32: 169-174.
13. Hughes TF, Borenstein AR, Schofield E, Wu Y, Larson EB, (2009) Association between late-life body mass index and dementia: The Kame Project. Neurology 72: 1741-1746.
14. Forti P, Pisacane N, Rietti E, Lucicesare A, Olivelli V, et al. (2010) Metabolic syndrome and risk of dementia in older adults. J Am Geriatr Soc 58: 487-492.
15. Gunstad J, Paul RH, Brickman AM, Cohen RA, Arns M, et al. (2006) Patterns of cognitive performance in middle-aged and older adults: A cluster analytic examination. J Geriatr Psychiatry Neurol 19: 59-64.
16. Gunstad J, Paul RH, Cohen RA, Tate DF, Spitznagel MB, et al. (2007) Elevated body mass index is associated with executive dysfunction in otherwise healthy adults. Compr Psychiatry 48: 57-61.
17. Jeong SK, Nam HS, Son MH, Son EJ, Cho KH, (2005) Interactive effect of obesity indexes on cognition. Dement Geriatr Cogn Disord 19: 91-96.
18. Gunstad J, Paul RH, Cohen RA, Tate DF, Spitznagel MB, et al. (2008) Relationship between body mass index and brain volume in healthy adults. Int J Neurosci 118: 1582-1593.
19. LeBlanc AC, Chen HY, Autilio-Gambetti L, Gambetti P, (1991) Differential APP gene expression in rat cerebral cortex, meninges, and primary astroglial, microglial and neuronal cultures. FEBS Lett 292: 171-178.
20. Kang J, Lemaire HG, Unterbeck A, Salbaum JM, Masters CL, et al. (1987) The precursor of Alzheimer's disease amyloid A4 protein resembles a cell-surface receptor. Nature 325: 733-736.
21. Goate A, Chartier-Harlin MC, Mullan M, Brown J, Crawford F, et al. (1991) Segregation of a missense mutation in the amyloid precursor protein gene with familial Alzheimer's disease. Nature 349: 704-706.
22. Schellenberg GD, Bird TD, Wijsman EM, Orr HT, Anderson L, et al. (1992) Genetic linkage evidence for a familial Alzheimer's disease locus on chromosome 14. Science (New York, NY) 258: 668-671.
23. Sherrington R, Rogaev EI, Liang Y, Rogaeva EA, Levesque G, et al. (1995) Cloning of a gene bearing missense mutations in early-onset familial Alzheimer's disease. Nature 375: 754-760.
24. Levy-Lahad E, Wasco W, Poorkaj P, Romano DM, Oshima J, et al. (1995) Candidate gene for the chromosome 1 familial Alzheimer's disease locus. Science (New York, NY) 269: 973-977.
25. Lee YH, Tharp WG, Maple RL, Nair S, Permana PA, et al. (2008) Amyloid precursor protein expression is upregulated in adipocytes in obesity. Obesity (Silver Spring, Md) 16: 1493-1500.
26. Lee YH, Martin JM, Maple RL, Tharp WG, Pratley RE, (2009) Plasma amyloid-beta peptide levels correlate with adipocyte amyloid precursor protein gene expression in obese individuals. Neuroendocrinology 90: 383-390.
27. Sommer G, Kralisch S, Lipfert J, Weise S, Krause K, et al. (2009) Amyloid precursor protein expression is induced by tumor necrosis factor alpha in 3T3-L1 adipocytes. J Cell Biochem 108: 1418-1422.
28. Balakrishnan K, Verdile G, Mehta PD, Beilby J, Nolan D, et al. (2005) Plasma Abeta42 correlates positively with increased body fat in healthy individuals. J Alzheimers Dis 8: 269-282.
29. Leahey TM, Myers TA, Gunstad J, Glickman E, Spitznagel MB, et al. (2007) Abeta40 is associated with cognitive function, body fat and physical fitness in healthy older adults. Nutr Neurosci 10: 205-209.
30. Levin-Allerhand JA, Lominska CE, Smith JD, (2002) Increased amyloid- levels in APPSWE transgenic mice treated chronically with a physiological high-fat high-cholesterol diet. J Nutr Health Aging 6: 315-319.
31. Kohjima M, Sun Y, Chan L, (2010) Increased food intake leads to obesity and insulin resistance in the tg2576 Alzheimer's disease mouse model. Endocrinology 151: 1532-1540.
32. Thirumangalakudi L, Prakasam A, Zhang R, Bimonte-Nelson H, Sambamurti K, et al. (2008) High cholesterol-induced neuroinflammation and amyloid precursor protein processing correlate with loss of working memory in mice. Journal of neurochemistry 106: 475-485.
33. Lang E, Szendrei GI, Lee VM, Otvos L Jr, (1992) Immunological and conformation characterization of a phosphorylated immunodominant epitope on the paired helical filaments found in Alzheimer's disease. Biochem Biophys Res Commun 187: 783-790.
34. Bendotti C, Forloni GL, Morgan RA, O'Hara BF, Oster-Granite ML, et al. (1988) Neuroanatomical localization and quantification of amyloid precursor protein mRNA by in situ hybridization in the brains of normal, aneuploid, and lesioned mice. Proceedings of the National Academy of Sciences of the United States of America 85: 3628-3632.
35. Floden AM, Combs CK, (2006) Beta-amyloid stimulates murine postnatal and adult microglia cultures in a unique manner. J Neurosci 26: 4644-4648.
36. Borst SE, Conover CF, (2005) High-fat diet induces increased tissue expression of TNF-alpha. Life Sciences 77: 2156-2165.
37. Ibrahim MM, (2010) Subcutaneous and visceral adipose tissue: structural and functional differences. Obesity reviews: an official journal of the International Association for the Study of Obesity 11: 11-18.
38. Matsubara Y, Kano K, Kondo D, Mugishima H, Matsumoto T, (2009) Differences in adipocytokines and fatty acid composition between two adipocyte fractions of small and large cells in high-fat diet-induced obese mice. Annals of Nutrition & Metabolism 54: 258-267.
39. Poussin C, Hall D, Minehira K, Galzin AM, Tarussio D, et al. (2008) Different transcriptional control of metabolism and extracellular matrix in visceral and subcutaneous fat of obese and rimonabant treated mice. PloS one 3: e3385.
40. Hotamisligil GS, Arner P, Caro JF, Atkinson RL, Spiegelman BM, (1995) Increased adipose tissue expression of tumor necrosis factor-alpha in human obesity and insulin resistance. The Journal of clinical investigation 95: 2409-2415.
41. Kern PA, Saghizadeh M, Ong JM, Bosch RJ, Deem R, et al. (1995) The expression of tumor necrosis factor in human adipose tissue. Regulation by obesity, weight loss, and relationship to lipoprotein lipase. The Journal of clinical investigation 95: 2111-2119.
42. Hotamisligil GS, Shargill NS, Spiegelman BM, (1993) Adipose expression of tumor necrosis factor-alpha: direct role in obesity-linked insulin resistance. Science (New York, NY) 259: 87-91.
43. Sondag CM, Combs CK, (2004) Amyloid precursor protein mediates proinflammatory activation of monocytic lineage cells. The Journal of biological chemistry 279: 14456-14463.
44. Sondag CM, Combs CK, (2006) Amyloid precursor protein cross-linking stimulates beta amyloid production and pro-inflammatory cytokine release in monocytic lineage cells. Journal of neurochemistry 97: 449-461.
45. Sondag CM, Combs CK, (2010) Adhesion of monocytes to type I collagen stimulates an APP-dependent proinflammatory signaling response and release of Abeta1-40. Journal of neuroinflammation 7: 22.
46. Weisberg SP, McCann D, Desai M, Rosenbaum M, Leibel RL, et al. (2003) Obesity is associated with macrophage accumulation in adipose tissue. The Journal of clinical investigation 112: 1796-1808.
47. Kanda H, Tateya S, Tamori Y, Kotani K, Hiasa K, et al. (2006) MCP-1 contributes to macrophage infiltration into adipose tissue, insulin resistance, and hepatic steatosis in obesity. The Journal of clinical investigation 116: 1494-1505.
48. Coenen KR, Gruen ML, Chait A, Hasty AH, (2007) Diet-induced increases in adiposity, but not plasma lipids, promote macrophage infiltration into white adipose tissue. Diabetes 56: 564-573.
49. Burgess AW, Camakaris J, Metcalf D, (1977) Purification and properties of colony-stimulating factor from mouse lung-conditioned medium. J Biol Chem 252: 1998-2003.
50. Wallet MA, Wallet SM, Guiulfo G, Sleasman JW, Goodenow MM, (2010) IFNgamma primes macrophages for inflammatory activation by high molecular weight hyaluronan. Cell Immunol 262: 84-88.
51. Schroder K, Sweet MJ, Hume DA, (2006) Signal integration between IFNgamma and TLR signalling pathways in macrophages. Immunobiology 211: 511-524.
52. Doherty TM, Kastelein R, Menon S, Andrade S, Coffman RL, (1993) Modulation of murine macrophage function by IL-13. J Immunol 151: 7151-7160.
53. Lee YH, Pratley RE, (2007) Abdominal obesity and cardiovascular disease risk: the emerging role of the adipocyte. J Cardiopulm Rehabil Prev 27: 2-10.
54. Ho L, Qin W, Pompl PN, Xiang Z, Wang J, et al. (2004) Diet-induced insulin resistance promotes amyloidosis in a transgenic mouse model of Alzheimer's disease. Faseb J 18: 902-904.
55. Moroz N, Tong M, Longato L, Xu H, de la Monte SM, (2008) Limited Alzheimer-type neurodegeneration in experimental obesity and type 2 diabetes mellitus. J Alzheimers Dis 15: 29-44.
56. van Dielen FM, van't Veer C, Schols AM, Soeters PB, Buurman WA, et al. (2001) Increased leptin concentrations correlate with increased concentrations of inflammatory markers in morbidly obese individuals. Int J Obes Relat Metab Disord 25: 1759-1766.
57. Poitou C, Coussieu C, Rouault C, Coupaye M, Cancello R, et al. (2006) Serum amyloid A: a marker of adiposity-induced low-grade inflammation but not of metabolic status. Obesity (Silver Spring) 14: 309-318.
58. Fain JN, (2010) Release of inflammatory mediators by human adipose tissue is enhanced in obesity and primarily by the nonfat cells: a review. Mediators Inflamm 2010.
59. Antuna-Puente B, Feve B, Fellahi S, Bastard JP, (2008) Adipokines: the missing link between insulin resistance and obesity. Diabetes Metab 34: 2-11.
60. Fain JN, (2006) Release of interleukins and other inflammatory cytokines by human adipose tissue is enhanced in obesity and primarily due to the nonfat cells. Vitam Horm 74: 443-477.
61. Fain JN, Tagele BM, Cheema P, Madan AK, Tichansky DS, (2010) Release of 12 adipokines by adipose tissue, nonfat cells, and fat cells from obese women. Obesity (Silver Spring) 18: 890-896.
62. Schmechel DE, Goldgaber D, Burkhart DS, Gilbert JR, Gajdusek DC, et al. (1988) Cellular localization of messenger RNA encoding amyloid-beta-protein in normal tissue and in Alzheimer disease. Alzheimer Dis Assoc Disord 2: 96-111.
63. Bauer J, Konig G, Strauss S, Jonas U, Ganter U, et al. (1991) In-vitro matured human macrophages express Alzheimer's beta A4-amyloid precursor protein indicating synthesis in microglial cells. FEBS Lett 282: 335-340.
64. Haass C, Hung AY, Selkoe DJ, (1991) Processing of beta-amyloid precursor protein in microglia and astrocytes favors an internal localization over constitutive secretion. J Neurosci 11: 3783-3793.
65. Banati RB, Gehrmann J, Czech C, Monning U, Jones LL, et al. (1993) Early and rapid de novo synthesis of Alzheimer beta A4-amyloid precursor protein (APP) in activated microglia. Glia 9: 199-210.
66. Banati RB, Gehrmann J, Kreutzberg GW, (1994) Glial beta-amyloid precursor protein: expression in the dentate gyrus after entorhinal cortex lesion. Neuroreport 5: 1359-1361.
67. Banati RB, Gehrmann J, Monning U, Czech C, Beyreuther K, et al. (1994) Amyloid precursor protein (APP) as a microglial acute phase protein. Neuropathol Appl Neurobiol 20: 194-195.
68. Banati RB, Gehrmann J, Lannes-Vieira J, Wekerle H, Kreutzberg GW, (1995) Inflammatory reaction in experimental autoimmune encephalomyelitis (EAE) is accompanied by a microglial expression of the beta A4-amyloid precursor protein (APP). Glia 14: 209-215.
69. Banati RB, Gehrmann J, Wiessner C, Hossmann KA, Kreutzberg GW, (1995) Glial expression of the beta-amyloid precursor protein (APP) in global ischemia. J Cereb Blood Flow Metab 15: 647-654.
70. Gehrmann J, Banati RB, Cuzner ML, Kreutzberg GW, Newcombe J, (1995) Amyloid precursor protein (APP) expression in multiple sclerosis lesions. Glia 15: 141-151.
71. Gehrmann J, Banati RB, Wiessner C, Hossmann KA, Kreutzberg GW, (1995) Reactive microglia in cerebral ischaemia: an early mediator of tissue damage? Neuropathol Appl Neurobiol 21: 277-289.
72. Monning U, Sandbrink R, Banati RB, Masters CL, Beyreuther K, (1994) Transforming growth factor beta mediates increase of mature transmembrane amyloid precursor protein in microglial cells. FEBS Lett 342: 267-272.
73. Monning U, Sandbrink R, Weidemann A, Banati RB, Masters CL, et al. (1995) Extracellular matrix influences the biogenesis of amyloid precursor protein in microglial cells. J Biol Chem 270: 7104-7110.
74. Blume AJ, Vitek MP, (1989) Focusing on IL-1-promotion of beta-amyloid precursor protein synthesis as an early event in Alzheimer's disease. Neurobiol Aging 10: 406-408; discussion 412-404.
75. Forloni G, Demicheli F, Giorgi S, Bendotti C, Angeretti N, (1992) Expression of amyloid precursor protein mRNAs in endothelial, neuronal and glial cells: modulation by interleukin-1. Brain Res Mol Brain Res 16: 128-134.
76. Hung AY, Koo EH, Haass C, Selkoe DJ, (1992) Increased expression of beta-amyloid precursor protein during neuronal differentiation is not accompanied by secretory cleavage. Proc Natl Acad Sci U S A 89: 9439-9443.
77. Ohyagi Y, Tabira T, (1993) Effect of growth factors and cytokines on expression of amyloid beta protein precursor mRNAs in cultured neural cells. Brain Res Mol Brain Res 18: 127-132.
78. Smith CJ, Johnson EM Jr, Osborne P, Freeman RS, Neveu I, et al. (1993) NGF deprivation and neuronal degeneration trigger altered beta-amyloid precursor protein gene expression in the rat superior cervical ganglia in vivo and in vitro. Brain Res Mol Brain Res 17: 328-334.
79. Sola C, Garcia-Ladona FJ, Mengod G, Probst A, Frey P, et al. (1993) Increased levels of the Kunitz protease inhibitor-containing beta APP mRNAs in rat brain following neurotoxic damage. Brain Res Mol Brain Res 17: 41-52.
80. Willoughby DA, Johnson SA, Pasinetti GM, Tocco G, Najm I, et al. (1992) Amyloid precursor protein mRNA encoding the Kunitz protease inhibitor domain is increased by kainic acid-induced seizures in rat hippocampus. Exp Neurol 118: 332-339.
81. Meda L, Cassatella MA, Szendrei GI, Otvos L Jr, Baron P, et al. (1995) Activation of microglial cells by beta-amyloid protein and interferon-gamma. Nature 374: 647-650.
82. Bitting L, Naidu A, Cordell B, Murphy GM Jr, (1996) Beta-amyloid peptide secretion by a microglial cell line is induced by beta-amyloid-(25-35) and lipopolysaccharide. J Biol Chem 271: 16084-16089.
83. Lorton D, Kocsis JM, King L, Madden K, Brunden KR, (1996) beta-Amyloid induces increased release of interleukin-1 beta from lipopolysaccharide-activated human monocytes. J Neuroimmunol 67: 21-29.
84. Lorton D, Schaller J, Lala A, De Nardin E, (2000) Chemotactic-like receptors and Abeta peptide induced responses in Alzheimer's disease. Neurobiol Aging 21: 463-473.
85. Lue LF, Walker DG, Brachova L, Beach TG, Rogers J, et al. (2001) Involvement of microglial receptor for advanced glycation endproducts (RAGE) in Alzheimer's disease: identification of a cellular activation mechanism. Exp Neurol 171: 29-45.
86. Combs CK, Johnson DE, Cannady SB, Lehman TM, Landreth GE, (1999) Identification of microglial signal transduction pathways mediating a neurotoxic response to amyloidogenic fragments of beta-amyloid and prion proteins. J Neurosci 19: 928-939.
87. Sondag CM, Dhawan G, Combs CK, (2009) Beta amyloid oligomers and fibrils stimulate differential activation of primary microglia. J Neuroinflammation 6: 1.
88. Wu SZ, Bodles AM, Porter MM, Griffin WS, Basile AS, et al. (2004) Induction of serine racemase expression and D-serine release from microglia by amyloid beta-peptide. J Neuroinflammation 1: 2.
89. Hayden KM, Zandi PP, Khachaturian AS, Szekely CA, Fotuhi M, et al. (2007) Does NSAID use modify cognitive trajectories in the elderly? The Cache County study. Neurology 69: 275-282.
90. Wang J, Ho L, Qin W, Rocher AB, Seror I, et al. (2005) Caloric restriction attenuates beta-amyloid neuropathology in a mouse model of Alzheimer's disease. Faseb J 19: 659-661.
91. Kim DH, Sandoval D, Reed JA, Matter EK, Tolod EG, et al. (2008) The role of GM-CSF in adipose tissue inflammation. Am J Physiol Endocrinol Metab 295: E1038-1046.
92. McGillicuddy FC, Chiquoine EH, Hinkle CC, Kim RJ, Shah R, et al. (2009) Interferon gamma attenuates insulin signaling, lipid storage, and differentiation in human adipocytes via activation of the JAK/STAT pathway. J Biol Chem 284: 31936-31944.
93. O'Rourke RW, Metcalf MD, White AE, Madala A, Winters BR, et al. (2009) Depot-specific differences in inflammatory mediators and a role for NK cells and IFN-gamma in inflammation in human adipose tissue. Int J Obes (Lond) 33: 978-990.
94. Rocha VZ, Folco EJ, Sukhova G, Shimizu K, Gotsman I, et al. (2008) Interferon-gamma, a Th1 cytokine, regulates fat inflammation: a role for adaptive immunity in obesity. Circ Res 103: 467-476.
95. Shaul ME, Bennett G, Strissel KJ, Greenberg AS, Obin MS, (2010) Dynamic, M2-like remodeling phenotypes of CD11c+ adipose tissue macrophages during high-fat diet--induced obesity in mice. Diabetes 59: 1171-1181.
96. Meijer K, de Vries M, Al-Lahham S, Bruinenberg M, Weening D, et al. (2011) Human primary adipocytes exhibit immune cell function: adipocytes prime inflammation independent of macrophages. PLoS One 6: e17154.
97. Nakarai H, Yamashita A, Nagayasu S, Iwashita M, Kumamoto S, et al. (2011) Adipocyte-macrophage interaction may mediate LPS-induced low-grade inflammation: potential link with metabolic complications. Innate Immun.
98. Gomez-Casanovas E, Sanmarti R, Sole M, Canete JD, Munoz-Gomez J, (2001) The clinical significance of amyloid fat deposits in rheumatoid arthritis: a systematic long-term followup study using abdominal fat aspiration. Arthritis Rheum 44: 66-72.
99. Barile L, Ariza R, Muci H, Pizarro S, Fraga A, et al. (1993) Tru-cut needle biopsy of subcutaneous fat in the diagnosis of secondary amyloidosis in rheumatoid arthritis. Arch Med Res 24: 189-192.
100. Bradford MM, (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72: 248-254.
101. Golovko MY, Murphy EJ, (2008) An improved LC-MS/MS procedure for brain prostanoid analysis using brain fixation with head-focused microwave irradiation and liquid-liquid extraction. J Lipid Res 49: 893-902.
102. Fernyhough ME, Vierck JL, Hausman GJ, Mir PS, Okine EK, et al. (2004) Primary adipocyte culture: adipocyte purification methods may lead to a new understanding of adipose tissue growth and development. Cytotechnology 46: 163-172.