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Current Neuropharmacology


ISSN (Print): 1570-159X
ISSN (Online): 1875-6190

Systematic Review Article

Using Cell Cultures for the Investigation of Treatments for Attention Deficit Hyperactivity Disorder: A Systematic Review

Author(s): Danielly Chierrito, Camila B. Villas-Boas, Fernanda S. Tonin, Fernando Fernandez-Llimos, Andréia C.C. Sanches and João C.P. de Mello*

Volume 17 , Issue 10 , 2019

Page: [916 - 925] Pages: 10

DOI: 10.2174/1570159X17666190409143155

Price: $65


Background: Advances in basic and molecular biology have promoted the use of cell cultures in a wide range of areas, including the evaluation of drug efficacy, safety and toxicity.

Objective: This article aims to provide a general overview of the methodological parameters of cell cultures used to investigate therapeutic options for Attention Deficit Hyperactivity Disorder.

Method: A systematic search was performed in the electronic databases PubMed, Scopus, and DOAJ. In vitro experimental studies using cell cultures were included.

Results: A total of 328 studies were initially identified, with 16 included for qualitative synthesis. Seven studies used neuronal cells (SH-SY5Y neuroblastoma and PC12 cell line) and nine used nonneuronal cells. All the studies described the culture conditions, but most studies were inconsistent with regard to reporting results and raw data. Only one-third of the studies performed cell viability assays, while a further 30% conducted gene expression analysis. Other additional tests included electrophysiological evaluation and transporter activity. More than 50% of the studies evaluated the effects of drugs such as methylphenidate and atomoxetine, while plant extracts were assessed in four studies and polyunsaturated fatty acids in one.

Conclusion: We suggested a flowchart to guide the planning and execution of studies, and a checklist to be completed by authors to allow the standardized reporting of results. This may guide the elaboration of laboratory protocols and further in vitro studies.

Keywords: Cell model, neuronal cell, ADHD, ADHD treatment, methodological aspect, flowchart, checklist.

Graphical Abstract
Yao, T.; Asayama, Y. Animal-cell culture media: History, characteristics, and current issues. Reprod. Med. Biol., 2017, 16(2), 99-117.
[] [PMID: 29259457]
Carter, M.; Shieh, J.C. Guide to Research Techniques in Neuroscience; Cell Culture Techniques, 2010, pp. 281-296.
Simon, V.; Czobor, P.; Bálint, S.; Mészáros, A.; Bitter, I. Prevalence and correlates of adult attention-deficit hyperactivity disorder: meta-analysis. Br. J. Psychiatry, 2009, 194(3), 204-211.
[] [PMID: 19252145]
American Psychiatry Association (APA). Diagnostic and statistical manual of mental disorders., Fifth Edition. American Psychiatric Publishing., (5th ed. ). 2013.
Polanczyk, G.V.; Salum, G.A.; Sugaya, L.S.; Caye, A.; Rohde, L.A. Annual research review: A meta-analysis of the worldwide prevalence of mental disorders in children and adolescents. J. Child Psychol. Psychiatry, 2015, 56(3), 345-365.
[ 10.1111/jcpp.12381] [PMID: 25649325]
Thomas, R.; Sanders, S.; Doust, J.; Beller, E.; Glasziou, P. Prevalence of attention-deficit/hyperactivity disorder: a systematic review and meta-analysis. Pediatrics, 2015, 135(4), e994-e1001.
[] [PMID: 25733754]
Schmidt, A.J.; Krieg, J.C.; Clement, H.W.; Gebhardt, S.; Schulz, E.; Heiser, P. Impact of drugs approved for treating ADHD on the cell survival and energy metabolism: an in-vitro study in human neuronal and immune cells. J. Psychopharmacol. (Oxford), 2010, 24(12), 1829-1833.
[] [PMID: 19605603]
Schmidt, A.J.; Krieg, J-C.; Hemmeter, U.M.; Kircher, T.; Schulz, E.; Clement, H-W.; Heiser, P. Impact of plant extracts tested in attention-deficit/hyperactivity disorder treatment on cell survival and energy metabolism in human neuroblastoma SH-SY5Y cells. Phytother. Res., 2010, 24(10), 1549-1553 b.
[] [PMID: 20878709]]
Knörle, R. Extracts of Sideritis scardica as triple monoamine reuptake inhibitors. J. Neural Transm. (Vienna), 2012, 119(12), 1477-1482.
[] [PMID: 22622367]
Feio-Azevedo, R.; Costa, V.M.; Ferreira, L.M.; Branco, P.S.; Pereira, F.C.; Bastos, M.L.; Carvalho, F.; Capela, J.P. Toxicity of the amphetamine metabolites 4-hydroxyamphetamine and 4-hydroxynorephedrine in human dopaminergic differentiated SH-SY5Y cells. Toxicol. Lett., 2017, 269, 65-76.
[ 10.1016/j.toxlet.2017.01.012] [PMID: 28115274]
Thapar, A.; Cooper, M. Attention deficit hyperactivity disorder. Lancet, 2016, 387(10024), 1240-1250.
[ S0140-6736(15)00238-X] [PMID: 26386541]
Li, Y.; Gao, J.; He, S.; Zhang, Y.; Wang, Q. An evaluation on the efficacy and safety of treatments for Attention Deficit Hyperactivity Disorder in children and adolescents: a comparison of multiple treatments. Mol. Neurobiol., 2017, 54(9), 6655-6669.
[] [PMID: 27738872]
Van Craenenbroeck, K.; Gellynck, E.; Lintermans, B.; Leysen, J.E.; Van Tol, H.H.; Haegeman, G.; Vanhoenacker, P. Influence of the antipsychotic drug pipamperone on the expression of the dopamine D4 receptor. Life Sci., 2006, 80(1), 74-81.
[] [PMID: 16978659]
Suter, W.; Martus, H-J.; Elhajouji, A. Methylphenidate is not clastogenic in cultured human lymphocytes and in the mouse bone-marrow micronucleus test. Mutat. Res., 2006, 607(2), 153-159.
[] [PMID: 16829163]
Zhao, G.; Li, S.; Qin, G-W.; Fei, J.; Guo, L-H. Inhibitive effects of Fructus Psoraleae extract on dopamine transporter and noradrenaline transporter. J. Ethnopharmacol., 2007, 112(3), 498-506.
[] [PMID: 17555897]
Wakamatsu, A.; Nomura, S.; Tate, Y.; Shimizu, S.; Harada, Y. Effects of methylphenidate hydrochloride on the cardiovascular system in vivo and in vitro: a safety pharmacology study. J. Pharmacol. Toxicol. Methods, 2009, 59(3), 128-134.
[] [PMID: 19281853]
Ludolph, A.G.; Udvardi, P.T.; Schaz, U.; Henes, C.; Adolph, O.; Weigt, H.U.; Fegert, J.M.; Boeckers, T.M.; Föhr, K.J. Atomoxetine acts as an NMDA receptor blocker in clinically relevant concentrations. Br. J. Pharmacol., 2010, 160(2), 283-291.
[] [PMID: 20423340]
Bartl, J.; Walitza, S.; Grünblatt, E. Enhancement of cell viability after treatment with polyunsaturated fatty acids. Neurosci. Lett., 2014, 559, 56-60.
[] [PMID: 24269370]
Schwarz, R.; Reif, A.; Scholz, C-J.; Weissflog, L.; Schmidt, B.; Lesch, K-P.; Jacob, C.; Reichert, S.; Heupel, J.; Volkert, J.; Kopf, J.; Hilscher, M.; Weber, H.; Kittel-Schneider, S. A preliminary study on methylphenidate-regulated gene expression in lymphoblastoid cells of ADHD patients. World J. Biol. Psychiatry, 2015, 16(3), 180-189.
[ 948064] [PMID: 25162476]
Kittel-Schneider, S.; Spiegel, S.; Renner, T.; Romanos, M.; Reif, A.; Reichert, S.; Heupel, J.; Schnetzler, L.; Stopper, H.; Jacob, C. Cytogenetic effects of chronic methylphenidate treatment and chronic social stress in adults with Attention-Deficit/Hyperactivity Disorder. Pharmacopsychiatry, 2016, 49(4), 146-154.
[] [PMID: 26926233]
Salviano, L.H.M.S.; Linhares, M.I.; de Lima, K.A.; de Souza, A.G.; Lima, D.B.; Jorge, A.R.C.; da Costa, M.F.; Filho, A.J.M.C.; Martins, A.M.C.; Monteiro, H.S.A.; de Jesus Ponte Carvalho, T.M.; de França Fonteles, M.M. Study of the safety of methylphenidate: Focus on nephrotoxicity aspects. Life Sci., 2015, 141, 137-142.
[] [PMID: 26407472]
Larson, D.R.; Singer, R.H.; Zenklusen, D. A single molecule view of gene expression. Trends Cell Biol., 2009, 19(11), 630-637.
[] [PMID: 19819144]
Xie, H.R.; Hu, L.S.; Li, G.Y. SH-SY5Y human neuroblastoma cell line: in vitro cell model of dopaminergic neurons in Parkinson’s disease. Chin. Med. J. (Engl.), 2010, 123(8), 1086-1092.
[PMID: 20497720]
Adan, A.; Kiraz, Y.; Baran, Y. Cell proliferation and cytotoxicity assays. Curr. Pharm. Biotechnol., 2016, 17(14), 1213-1221.
[] [PMID: 27604355]
Viswanath, B.; Jose, S.P.; Squassina, A.; Thirthalli, J.; Purushottam, M.; Mukherjee, O.; Vladimirov, V.; Patrinos, G.P.; Del Zompo, M.; Jain, S. Cellular models to study bipolar disorder: A systematic review. J. Affect. Disord., 2015, 184, 36-50.
[] [PMID: 26070045]
Xicoy, H.; Wieringa, B.; Martens, G.J.M. The SH-SY5Y cell line in Parkinson’s disease research: a systematic review. Mol. Neurodegener., 2017, 12(1), 10.
[] [PMID: 28118852]
Liberati, A.; Altman, D.G.; Tetzlaff, J.; Mulrow, C.; Gotzsche, P.C.; Ioannidis, J.P.A.; Clarke, M.; Devereaux, P.J.; Kleijnen, J.; Moher, D. The PRISMA statement for reporting systematic reviews and meta-analyses of studies that evaluate health care interventions: Explanation and elaboration. BMJ, 2007, 339.
Higgins, J.P.T.; Green, S. Cochrane handbook for systematic reviews of interventions The Cochrane Collaboration., 2010.Available at:
Joanna Briggs Institute. Methodology for JBI scoping reviews Joanna Briggs Institute. Joanna Briggs Institute Reviewers’ Manual:2015 edition., Available at: docs/sumari/Reviewers-Manual_Methodology-for-JBI-Scoping-Reviews_2015_v2.pdf2018.
Hooijmans, C.R.; Rovers, M.M.; de Vries, R.B.; Leenaars, M.; Ritskes-Hoitinga, M.; Langendam, M.W. SYRCLE’s risk of bias tool for animal studies. BMC Med. Res. Methodol., 2014, 14, 43.
[] [PMID: 24667063]
Bartl, J.; Link, P.; Schlosser, C.; Gerlach, M.; Schmitt, A.; Walitza, S.; Riederer, P.; Grünblatt, E. Effects of methylphenidate: the cellular point of view. Atten. Defic. Hyperact. Disord., 2010, 2(4), 225-232.
[] [PMID: 21432609]
Schmidt, A.J.; Clement, H-W.; Gebhardt, S.; Hemmeter, U.M.; Schulz, E.; Krieg, J-C.; Kircher, T.; Heiser, P. Impact of psychostimulants and atomoxetine on the expression of 8-hydroxyguanine glycosylase 1 in human cells. J. Neural Transm. (Vienna), 2010, 117(6), 793-797.
[] [PMID: 20467764]
Nam, Y.; Shin, E-J.; Shin, S.W.; Lim, Y.K.; Jung, J.H.; Lee, J.H.; Ha, J.R.; Chae, J.S.; Ko, S.K.; Jeong, J.H.; Jang, C-G.; Kim, H-C. YY162 prevents ADHD-like behavioral side effects and cytotoxicity induced by Aroclor1254 via interactive signaling between antioxidant potential, BDNF/TrkB, DAT and NET. Food Chem. Toxicol., 2014, 65, 280-292.
[] [PMID: 24394491]
Biedler, J.L.; Helson, L.; Spengler, B.A. Morphology and growth, tumorigenicity, and cytogenetics of human neuroblastoma cells in continuous culture. Cancer Res., 1973, 33(11), 2643-2652.
[PMID: 4748425]
Shafer, T.J.; Atchison, W.D. Transmitter, ion channel and receptor properties of pheochromocytoma (PC12) cells: a model for neurotoxicological studies. Neurotoxicology, 1991, 12(3), 473-492.
[PMID: 1720882]
Shipley, M.M.; Mangold, C.A.; Szpara, M.L. Differentiation of the SH-SY5Y human neuroblastoma cell line. J. Vis. Exp., 2016, 108(108), 53193.
[PMID: 26967710]
Ferreira, P.S.; Nogueira, T.B.; Costa, V.M.; Branco, P.S.; Ferreira, L.M.; Fernandes, E.; Bastos, M.L.; Meisel, A.; Carvalho, F.; Capela, J.P. Neurotoxicity of “ecstasy” and its metabolites in human dopaminergic differentiated SH-SY5Y cells. Toxicol. Lett., 2013, 216(2-3), 159-170.
[ 11.015] [PMID: 23194825]
Bustin, S.A.; Nolan, T. Pitfalls of quantitative real-time reverse-transcription polymerase chain reaction. J. Biomol. Tech., 2004, 15(3), 155-166.
[PMID: 15331581]
Sharp, S.I.; McQuillin, A.; Gurling, H.M.D. Genetics of attention-deficit hyperactivity disorder (ADHD). Neuropharmacology, 2009, 57(7-8), 590-600.
[ 011] [PMID: 19715710]
Food and Drug Administration (FDA). S7B Nonclinical evaluation of the potential for delayed ventricular repolarization (QT interval prolongation) by Human Pharmaceuticals., Guidance for Industry. 2005.
Ioannidis, J.P. Why most published research findings are false. PLoS Med., 2005, 2(8)e124
[ 0020124] [PMID: 16060722]
Collins, F.S.; Tabak, L.A. Policy: NIH plans to enhance reproducibility. Nature, 2014, 505(7485), 612-613.
[ 1038/505612a] [PMID: 24482835]
Fiala, C.; Diamandis, E.P. How to reduce scientific irreproducibility: the 5-year reflection. Clin. Chem. Lab. Med., 2017, 55(12), 1845-1848.
[] [PMID: 29031017]
Begley, C.G.; Ioannidis, J.P. Reproducibility in science: improving the standard for basic and preclinical research. Circ. Res., 2015, 116(1), 116-126.
[ 303819] [PMID: 25552691]
Lieu, C.H.; Tan, A.C.; Leong, S.; Diamond, J.R.; Eckhardt, S.G. From bench to bedside: lessons learned in translating preclinical studies in cancer drug development. J. Natl. Cancer Inst., 2013, 105(19), 1441-1456.
[] [PMID: 24052618]
Wu, X.; Lin, M.; Li, Y.; Zhao, X.; Yan, F. Effects of DMEM and RPMI 1640 on the biological behavior of dog periosteum-derived cells. Cytotechnology, 2009, 59(2), 103-111.
[ 1007/s10616-009-9200-5] [PMID: 19496017]
Huang, Z.; Shao, W.; Gu, J.; Hu, X.; Shi, Y.; Xu, W.; Huang, C.; Lin, D. Effects of culture media on metabolic profiling of the human gastric cancer cell line SGC7901. Mol. Biosyst., 2015, 11(7), 1832-1840.
[] [PMID: 25925870]
Brunner, D.; Frank, J.; Appl, H.; Schöffl, H.; Pfaller, W.; Gstraunthaler, G. Serum-free cell culture: the serum-free media interactive online database. ALTEX, 2010, 27(1), 53-62.
[ 10.14573/altex.2010.1.53] [PMID: 20390239]
Perlman, D. Use of antibiotics in cell culture media. Methods Enzymol., 1979, 58, 110-116.
[ (79)58128-2] [PMID: 423753]
Mazzio, E.; Soliman, K.F.A. Pyruvic acid cytoprotection against 1-methyl-4-phenylpyridinium, 6-hydroxydopamine and hydrogen peroxide toxicities in vitro. Neurosci. Lett., 2003, 337(2), 77-80.
[] [PMID: 12527392]
Wang, X.; Perez, E.; Liu, R.; Yan, L-J.; Mallet, R.T.; Yang, S-H. Pyruvate protects mitochondria from oxidative stress in human neuroblastoma SK-N-SH cells. Brain Res., 2007, 1132(1), 1-9.
[] [PMID: 17174285]
Pasieka, A.E.; Morgan, J.F. Glutamine metabolism of normal and malignant cells cultivated in synthetic media. Nature, 1959, 183(4669), 1201-1202.
[] [PMID: 13657053]
Yang, H.; Wang, J.; Sun, J.; Liu, X.; Duan, W-M.; Qu, T. A new method to effectively and rapidly generate neurons from SH-SY5Y cells. Neurosci. Lett., 2016, 610(610), 43-47.
[ 10.1016/j.neulet.2015.10.047] [PMID: 26497914]
Bustin, S.A. Real-time reverse transcription PCR. Ncyclopedia of Diagnostic Genomics and Proteomics, 2005, 1131, 1135.
Bustin, S.A.; Benes, V.; Garson, J.A.; Hellemans, J.; Huggett, J.; Kubista, M.; Mueller, R.; Nolan, T.; Pfaffl, M.W.; Shipley, G.L.; Vandesompele, J.; Wittwer, C.T. The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clin. Chem., 2009, 55(4), 611-622.
[ 1373/clinchem.2008.112797] [PMID: 19246619]
Taylor, S.; Wakem, M.; Dijkman, G.; Alsarraj, M.; Nguyen, M. A practical approach to RT-qPCR-Publishing data that conform to the MIQE guidelines. Methods, 2010, 50(4), S1-S5.
[] [PMID: 20215014]
Chiarotto, A.; Ostelo, R.W.; Turk, D.C.; Buchbinder, R.; Boers, M. Core outcome sets for research and clinical practice. Braz. J. Phys. Ther., 2017, 21(2), 77-84.
[ 001] [PMID: 28460714]
Clarke, M.; Williamson, P.R. Core outcome sets and systematic reviews. Syst. Rev., 2016, 5, 11.
[] [PMID: 26792080]
Coecke, S.; Balls, M.; Bowe, G.; Davis, J.; Gstraunthaler, G.; Hartung, T.; Hay, R.; Merten, O-W.; Price, A.; Schechtman, L.; Stacey, G.; Stokes, W. Guidance on good cell culture practice. a report of the second ECVAM task force on good cell culture practice. Altern. Lab. Anim., 2005, 33(3), 261-287.
[PMID: 16180980]

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