Abstract
Original language | English |
---|---|
Journal | PLoS ONE |
Volume | 15 |
Issue number | 11 November |
DOIs | |
Publication status | Published - 2020 |
Keywords
- ELAV like protein 1
- protein kinase C beta
- protein kinase C beta II
- unclassified drug
- vasculotropin
- Article
- cardiovascular system
- CASMC cell line
- cell culture
- cell density
- cell function
- cell separation
- cell viability
- controlled study
- enzyme linked immunosorbent assay
- human
- human cell
- in vivo study
- molecular dynamics
- MTT assay
- nervous system
- neuroscience
- SH-SY5Y cell line
- signal transduction
- Western blotting
- biological model
- bioreactor
- cell communication
- cytology
- metabolism
- nerve cell
- smooth muscle cell
- tumor cell line
- Bioreactors
- Cell Communication
- Cell Line, Tumor
- Humans
- Models, Cardiovascular
- Models, Neurological
- Myocytes, Smooth Muscle
- Neurons
- Signal Transduction
Fingerprint
Dive into the research topics of 'Use of dual-flow bioreactor to develop a simplified model of nervous-cardiovascular systems crosstalk: A preliminary assessment'. Together they form a unique fingerprint.Cite this
- APA
- Standard
- Harvard
- Vancouver
- Author
- BIBTEX
- RIS
Use of dual-flow bioreactor to develop a simplified model of nervous-cardiovascular systems crosstalk: A preliminary assessment. / Marchesi, N.; Barbieri, A.; Fahmideh, F.; Govoni, S.; Ghidoni, A.; Parati, G.; Vanoli, E.; Pascale, A.; Calvillo, L.
In: PLoS ONE, Vol. 15, No. 11 November, 2020.Research output: Contribution to journal › Article › peer-review
}
TY - JOUR
T1 - Use of dual-flow bioreactor to develop a simplified model of nervous-cardiovascular systems crosstalk: A preliminary assessment
AU - Marchesi, N.
AU - Barbieri, A.
AU - Fahmideh, F.
AU - Govoni, S.
AU - Ghidoni, A.
AU - Parati, G.
AU - Vanoli, E.
AU - Pascale, A.
AU - Calvillo, L.
N1 - Export Date: 5 March 2021 CODEN: POLNC Correspondence Address: Pascale, A.; Department of Drug Sciences, Italy; email: alessia.pascale@unipv.it Correspondence Address: Calvillo, L.; Department of Cardiovascular, Italy; email: l.calvillo@auxologico.it Chemicals/CAS: vasculotropin, 127464-60-2 Funding text 1: This work was financially supported by Association I-CARE Europe Onlus. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Authors are grateful to Dr. Lidia Cova for the precious suggestions. References: Vozzi, F, Mazzei, D, Vinci, B, Vozzi, G, Sbrana, T, Ricotti, L, A flexible bioreactor system for constructing in vitro tissue and organ models (2011) Biotechnol Bioeng, 108 (9), pp. 2129-2140. , https://doi.org/10.1002/bit.23164, PMID: 21495015; Vandrangi, P, Sosa, M, Shyy, JYJ, Rodgers, VGJ., Flow-dependent mass transfer may trigger endothelial signaling cascades (2012) PLoS One, 7 (4), p. e35260. , https://doi.org/10.1371/journal.pone.0035260, PMID: 22558132; Mazzei, D, Guzzardi, MA, Giusti, S, Ahluwalia, A., A low shear stress modular bioreactor for connected cell culture under high flow rates (2010) Biotechnol Bioeng, 106 (1), pp. 127-137. , https://doi.org/10.1002/bit.22671, PMID: 20091740; Giusti, S, Sbrana, T, La Marca, M, Di Patria, V, Martinucci, V, Tirella, A, A novel dual-flow bioreactor simulates increased fluorescein permeability in epithelial tissue barriers (2014) Biotechnol J, 9 (9), pp. 1175-1184. , https://doi.org/10.1002/biot.201400004, PMID: 24756869; Iori, E, Vinci, B, Murphy, E, Marescotti, MC, Avogaro, A, Ahluwalia, A., Glucose and fatty acid metabolism in a 3 tissue in-vitro model challenged with normo- and hyperglycaemia (2012) PLoS One, 7 (4), pp. 1-9; Kovalevich, J, Langford, D., Considerations for the use of SH-SY5Y neuroblastoma cells in neurobiology (2013) Methods Mol Biol, 1078, pp. 9-21. , https://doi.org/10.1007/978-1-62703-640-5_2, PMID: 23975817; Govoni, S., Psychiatric and Neurological Effects of Cardiovascular Drugs (2020) Brain and Heart Dynamics, , https://doi.org/10.1007/978-3-319-90305-7_46-1, Govoni S., Politi P., Vanoli E. (eds) Springer, Cham; Parga, JA, Rodriguez-Perez, AI, Garcia-Garrote, M, Rodriguez-Pallares, J, Labandeira-Garcia, JL., Angiotensin II induces oxidative stress and upregulates neuroprotective signaling from the NRF2 and KLF9 pathway in dopaminergic cells (2018) Free Radic Biol Med, 129, pp. 394-406. , https://doi.org/10.1016/j.freeradbiomed.2018.10.409, PMID: 30315936; Kohno, M, Ohmori, K, Nozaki, S, Mizushige, K, Yasunari, K, Kano, H, Effects of valsartan on angiotensin II-induced migration of human coronary artery smooth muscle cells (2000) Hypertens Res, 23 (6), pp. 677-681. , https://doi.org/10.1291/hypres.23.677, PMID: 11131281; Govoni, S, Pascale, A, Amadio, M, Calvillo, L, D’Elia, E, Cereda, C, NGF and heart: Is there a role in heart disease? (2011) Pharmacol Res, 63 (4), pp. 266-277. , https://doi.org/10.1016/j.phrs.2010.12.017, PMID: 21195180; Calvillo, L., Gironacci, M.M., Crotti, L., Meroni, P.L., Parati, G., Neuroimmune crosstalk in the pathophysiology of hypertension (2019) Nat Rev Cardiol, 16, pp. 476-490. , https://doi.org/10.1038/s41569-019-0178-1, PMID: 30894678; Govoni, S., Politi, P., Vanoli, E., (2020) Edr. Brain and Heart Dynamics, , Springer ISBN: 978-3-030 28007–9; Calvillo, L, Vanoli, E, Andreoli, E, Besana, A, Omodeo, E, Gnecchi, M, Vagal Stimulation, Through its Nicotinic Action, Limits Infarct Size and the Inflammatory Response to Myocardial Ischemia and Reperfusion (2011) J Cardiovasc Pharmacol, 58 (5), pp. 500-507. , https://doi.org/10.1097/FJC.0b013e31822b7204, PMID: 21765369; Gemes, G., Rigaud, M., Dean, C., Hopp, F.A., Hogan, Q.H., Seagard, J., Baroreceptor Reflex is Suppressed in Rats that Develop Hyperalgesia Behavior after Nerve Injury (2009) Pain, 146 (3), pp. 293-300. , https://doi.org/10.1016/j.pain.2009.07.040, PMID: 19729245; Zubcevic, J, Santisteban, MM, Pitts, T, Baekey, DM, Perez, PD, Bolser, DC, Functional neural-bone marrow pathways: Implications in hypertension and cardiovascular disease (2014) Hypertension, 63 (6), pp. 129-140. , https://doi.org/10.1161/HYPERTENSIONAHA.114.02440, PMID: 24688127; Ahluwalia, A, Misto, A, Vozzi, F, Magliaro, C, Mattei, G, Marescotti, MC, Systemic and vascular inflammation in an in-vitro model of central obesity (2018) PLoS One, 13 (2), pp. 1-15; Abiodun, OA, Ola, MS., Role of brain renin angiotensin system in neurodegeneration: An update (2020) Saudi J Biol Sci, 27, pp. 905-912. , https://doi.org/10.1016/j.sjbs.2020.01.026, PMID: 32127770; Roger, VL., The heart-brain connection: From evidence to action (2017) Eur Heart J, 38, pp. 3229-3231. , https://doi.org/10.1093/eurheartj/ehx387, PMID: 29020365; Goldston, K., Baillie, AJ., Depression and coronary heart disease: a review of the epidemiological evidence, explanatory mechanisms and management approaches (2008) Clin Psychol Rev, 28, pp. 288-306. , https://doi.org/10.1016/j.cpr.2007.05.005, PMID: 17601644; Felder, R.B., Francis, J., Zhang, Z.H., Wei, S.G., Weiss, R.M., Johnson, AK, Heart failure and the brain: new perspectives (2003) Am J Physiol Regul Integr Comp Physiol, 284 (2), pp. R259-R276. , https://doi.org/10.1152/ajpregu.00317.2002, PMID: 12529279; Calvillo, L, Parati, G., Immune System and Mind-Body Medicine–An Overview (2020) Brain and Heart Dynamics, pp. 1-19. , (Govoni S, Politi P, Vanoli E. Eds) Springer ISBN: 978-3-030 28007–9; Uijl, E, Ren, L, Danser, AHJ., Angiotensin generation in the brain: A re-evaluation (2018) Clin Sci, 132, pp. 839-850. , https://doi.org/10.1042/CS20180236, PMID: 29712882; Huber, G, Schuster, F, Raasch, W., Brain renin-angiotensin system in the pathophysiology of cardiovascular diseases (2017) Pharmacol Res, 125, pp. 72-90. , https://doi.org/10.1016/j.phrs.2017.06.016, PMID: 28687340; Johns, EJ., Angiotensin II in the brain and the autonomic control of the kidney (2005) Exp Physiol, 90 (2), pp. 163-168. , https://doi.org/10.1113/expphysiol.2004.029025, PMID: 15604112; Bali, A, Jaggi, AS., Angiotensin II-triggered kinase signaling cascade in the central nervous system (2016) Rev Neurosci, 27 (3), pp. 301-315. , https://doi.org/10.1515/revneuro-2015-0041, PMID: 26574890; McKinley, MJ, Albiston, AL, Allen, AM, Mathai, ML, May, CN, McAllen, RM, The brain renin-angiotensin system: Location and physiological roles (2003) Int J Biochem Cell Biol, 35 (6), pp. 901-918. , https://doi.org/10.1016/s1357-2725(02)00306-0, PMID: 12676175; Grobe, JL, Xu, D, Sigmund, CD., An Intracellular Renin-Angiotensin System in Neurons: Fact, Hypothesis or Fantasy (2008) Physiol, 23, pp. 187-193. , https://doi.org/10.1152/physiol.00002.2008, PMID: 18697992; Xu, P, Sriramula, S, Lazartigues, E., ACE2/ANG-(1–7)/Mas pathway in the brain: The axis of good (2011) Am J Physiol—Regul Integr Comp Physiol, 300 (4), pp. 804-817. , https://doi.org/10.1152/ajpregu.00222.2010, PMID: 21178125; Lehn, A, Gelauff, J, Hoeritzauer, I, Ludwig, L, McWhirter, L, Williams, S, Functional neurological disorders: mechanisms and treatment (2016) J Neurol, 263 (3), pp. 611-620. , https://doi.org/10.1007/s00415-015-7893-2, PMID: 26410744; Särkämö, T, Sihvonen, AJ., Golden oldies and silver brains: Deficits, preservation, learning, and rehabilitation effects of music in ageing-related neurological disorders (2018) Cortex, 109, pp. 104-123. , https://doi.org/10.1016/j.cortex.2018.08.034, PMID: 30312779; Abbruzzese, G, Marchese, R, Avanzino, L, Pelosin, E., Rehabilitation for Parkinson’s disease: Current outlook and future challenges (2016) Park Relat Disord, 22, pp. S60-S64. , https://doi.org/10.1016/j.parkreldis.2015.09.005, Suppl. 1: PMID: 26360239; Winstein, CJ, Stein, J, Arena, R, Bates, B, Cherney, LR, Cramer, SC, Guidelines for Adult Stroke Rehabilitation and Recovery: A Guideline for Healthcare Professionals from the American Heart Association/American Stroke Association (2016) Stroke, 47 (6), pp. e98-169. , https://doi.org/10.1161/STR.0000000000000098, PMID: 27145936; Walter, A, Etienne-Selloum, N, Sarr, M, Kane, MO, Beretz, A S-KV., Angiotensin II induces the vascular expression of VEGF and MMP-2 in vivo: preventive effect of red wine polyphenols (2008) J Vasc Res, 45, pp. 386-394. , https://doi.org/10.1159/000121408, PMID: 18354258; Imanishi, T, Hano, T, Nishio, I., Angiotensin II potentiates vascular endothelial growth factor-induced proliferation and network formation of endothelial progenitor cells (2004) Hypertens Res, 27 (2), pp. 101-108. , https://doi.org/10.1291/hypres.27.101, PMID: 15005273; Doller, A, Schlepckow, K, Schwalbe, H, Pfeilschifter, J, Eberhardt, W., Tandem Phosphorylation of Serines 221 and 318 by Protein Kinase Cδ Coordinates mRNA Binding and Nucleocytoplasmic Shuttling of HuR (2010) Mol Cell Biol, 30 (6), pp. 1397-1410. , https://doi.org/10.1128/MCB.01373-09, PMID: 20086103; Parihar, SP, Ozturk, M, Marakalala, MJ, Loots, DT, Hurdayal, R, Maasdorp, DB, Protein kinase C-delta (PKCδ), a marker of inflammation and tuberculosis disease progression in humans, is important for optimal macrophage killing effector functions and survival in mice (2018) Mucosal Immunol, 11 (2), pp. 496-511. , https://doi.org/10.1038/mi.2017.68, PMID: 28832027; Amadio, M, Bucolo, C, Leggio, GM, Drago, F, Govoni, S, Pascale, A., The PKCβ/HuR/VEGF pathway in diabetic retinopathy (2010) Biochem Pharmacol, 80 (8), pp. 1230-1237. , https://doi.org/10.1016/j.bcp.2010.06.033, PMID: 20599775; Amadio, M, Scapagnini, G, Lupo, G, Drago, F, Govoni, S, Pascale, A., PKCβII/HuR/VEGF: A new molecular cascade in retinal pericytes for the regulation of VEGF gene expression (2008) Pharmacol Res, 57 (1), pp. 60-66. , https://doi.org/10.1016/j.phrs.2007.11.006, PMID: 18206386; Amadio, M, Osera, C, Lupo, G, Motta, C, Drago, F, Govoni, S, Protein kinase C activation affects, via the mRNA-binding Huantigen R/ELAV protein, vascular endothelial growth factor expression in a pericytic/endothelial coculture model (2012) Mol Vis, 18, pp. 2153-2164. , PMID: 22879735; Ahmed, S, Chauhan, VM, Ghaemmaghami, AM, Aylott, JW., New generation of bioreactors that advance extracellular matrix modelling and tissue engineering (2019) Biotechnol Lett, 41 (1), pp. 1-25. , https://doi.org/10.1007/s10529-018-2611-7, PMID: 30368691; Yu, Y, Shamsi, MH, Krastev, DL, Dryden, MD, Leung, Y WA., A microfluidic method for dopamine uptake measurements in dopaminergic neurons (2016) Lab Chip, 16, pp. 543-552. , https://doi.org/10.1039/c5lc01515d, PMID: 26725686; Edwards, M.E., Good, TA., Use of a mathematical model to estimate stress and strain during elevated pressure induced lamina cribrosa deformation (2001) Curr Eye Res, 23, pp. 215-225. , https://doi.org/10.1076/ceyr.23.3.215.5460, PMID: 11803484; Triyoso, DH, Good, TA., Pulsatile shear stress leads to DNA fragmentation in human SH-SY5Y neuroblastoma cell line (1999) J Physiol, 515 (2), pp. 355-365. , https://doi.org/10.1111/j.1469-7793.1999.355ac.x, PMID: 10050003; Liao, XH, Xiang, Y, Li, H, Zheng, DL, Xu, Y, Xi Yu, C, VEGF-A Stimulates STAT3 Activity via Nitrosylation of Myocardin to Regulate the Expression of Vascular Smooth Muscle Cell Differentiation Markers (2017) Sci Rep, 7 (1), pp. 1-11. , https://doi.org/10.1038/s41598-016-0028-x, PMID: 28127051; Ramakrishnan, S, Anand, V, Roy, S., Vascular endothelial growth factor signaling in hypoxia and inflammation (2014) J Neuroimmune Pharmacol, 9 (2), pp. 142-160. , https://doi.org/10.1007/s11481-014-9531-7, PMID: 24610033; Adair, TH, Gay, WJ MJ., Growth regulation of the vascular system: evidence for a metabolic hypothesis (1990) Am J Physiol, 259 (3), pp. R393-R404. , https://doi.org/10.1152/ajpregu.1990.259.3.R393, (Pt 2): PMID: 1697737; Prabhakaran, K, Sampson, DA, Hoehner, JC., Neuroblastoma survival and death: An in vitro model of hypoxia and metabolic stress (2004) J Surg Res, 116 (2), pp. 288-296. , https://doi.org/10.1016/j.jss.2003.08.008, PMID: 15013368; González, A, González-González, A, Alonso-González, C, Menéndez-Menéndez, J, Mart Ínez-Campa, C, Cos, S., Melatonin inhibits angiogenesis in SH-SY5Y human neuroblastoma cells by downregulation of VEGF (2017) Oncol Rep, 37 (4), pp. 2433-2440. , https://doi.org/10.3892/or.2017.5446, PMID: 28259965; Osada-Oka, M, Ikeda, T, Imaoka, S, Akiba, S, Sato, T., VEGF-enhanced proliferation under hypoxia by an autocrine mechanism in human vascular smooth muscle cells (2008) J Atheroscler Thromb, 15 (1), pp. 26-33. , https://doi.org/10.5551/jat.e533, PMID: 18270456; Miura, SI, Fujino, M, Matsuo, Y, Tanigawa, H, Saku, K., Nifedipine-induces vascular endothelial growth factor secretion from coronary smooth muscle cells promotes endothelial-containing receptor/fetal liver kinase-1/NO pathway (2005) Hypertens Res, 28 (2), pp. 147-153. , https://doi.org/10.1291/hypres.28.147, PMID: 16025742; Pascale, A, Govoni, S., The complex world of post-transcriptional mechanisms: Is their deregulation a common link for diseases? Focus on ELAV-like RNA-binding proteins (2012) Cell Mol Life Sci, 69 (4), pp. 501-517. , https://doi.org/10.1007/s00018-011-0810-7, PMID: 21909784; Wang, J, Zhou, X, Lu, H, Song, M, Zhao, J, Wang, Q., Fluoxetine induces vascular endothelial growth factor/Netrin over-expression via the mediation of hypoxia-inducible factor 1-alpha in SH-SY5Y cells (2016) J Neurochem, 136 (6), pp. 1186-1195. , https://doi.org/10.1111/jnc.13521, PMID: 26718749; Maugeri, G, D’Amico, AG, Rasà, DM, Saccone, S, Federico, C, Cavallaro, S, PACAP and VIP regulate hypoxia-inducible factors in neuroblastoma cells exposed to hypoxia (2018) Neuropeptides, 69, pp. 84-91. , https://doi.org/10.1016/j.npep.2018.04.009, PMID: 29699729; Nangaku, M, Inagi, R, Miyata, T, Fujita, T., Angiotensin-Induced Hypoxia in the Kidney: Functional and Structural Changes of the Renal Circulation Hypoxia and the Circulation, , (Roach RC, Wagner PD, Hackett PH, Editors), Springer US, Boston, MA, ISBN: 978–0; Wolf, G, Schroeder, R, Stahl, RAK., Angiotensin II induces hypoxia-inducible factor-1α in PC 12 cells through a posttranscriptional mechanism: Role of AT2 receptors (2004) Am J Nephrol, 24 (4), pp. 415-421. , https://doi.org/10.1159/000080086, PMID: 15308873; Jiang, L, Zhu, R, Bu, Q, Li, Y, Shao, X, Gu, H, Brain Renin–Angiotensin System Blockade Attenuates Methamphetamine-Induced Hyperlocomotion and Neurotoxicity (2018) Neurotherapeutics, 15 (2), pp. 500-510. , https://doi.org/10.1007/s13311-018-0613-8, PMID: 29464572; Huang, Y, Mao, Y, Li, H, Shen, G NG., Knockdown of Nrf2 inhibits angiogenesis by downregulating VEGF expression through PI3K/Akt signaling pathway in cerebral microvascular endothelial cells under hypoxic conditions (2018) Biochem Cell Biol, 96 (4), pp. 475-482. , https://doi.org/10.1139/bcb-2017-0291, PMID: 29373803; Zhao, Q, Ishibashi, M, Hiasa, K, Tan, C, Takeshita, A, Egashira, K., Essential Role of Vascular Endothelial Growth Factor in Angiotensin II–Induced Vascular Inflammation and Remodeling (2004) Hypertension, 44 (3), pp. 264-270. , https://doi.org/10.1161/01.HYP.0000138688.78906.6b, PMID: 15262905; Slone, S, Anthony, SR, Wu, X, Benoit, JB, Aube, J, Xu, L, Activation of HuR downstream of p38 MAPK promotes cardiomyocyte hypertrophy (2016) Cell Signal, 28 (11), pp. 1735-1741. , https://doi.org/10.1016/j.cellsig.2016.08.005, PMID: 27521603; Liu, C, Fan, Y, Zhou, L, Zhu, HY, Song, YC, Hu, L, Pretreatment of mesenchymal stem cells with angiotensin II enhances paracrine effects, angiogenesis, gap junction formation and therapeutic efficacy for myocardial infarction (2015) Int J Cardiol, 188 (1), pp. 22-32. , https://doi.org/10.1016/j.ijcard.2015.03.425, PMID: 25880576; Tamarat, R, Silvestre, JS, Kubis, N, Benessiano, J, Duriez, M, DeGasparo, M, Endothelial nitric oxide synthase lies downstream from angiotensin II-induced angiogenesis in ischemic hindlimb (2002) Hypertension, 39 (3), pp. 830-835. , https://doi.org/10.1161/hy0302.104671, PMID: 11897773; Tamarat, R, Silvestre, JS, Duriez, M, Levy, BI., Angiotensin II angiogenic effect in vivo involves vascular endothelial growth factor- and inflammation-related pathways (2002) Lab Investig, 82 (6), pp. 747-756. , https://doi.org/10.1097/01.lab.0000017372.76297.eb, PMID: 12065685; Ismadi, MZ, Gupta, P, Fouras, A, Verma, P, Jadhav, S, Bellare, J, Flow characterization of a spinner flask for induced pluripotent stem cell culture application (2014) PLoS One, 9 (10), p. e106493. , https://doi.org/10.1371/journal.pone.0106493, PMID: 25279733
PY - 2020
Y1 - 2020
N2 - Chronic conditions requiring long-term rehabilitation therapies, such as hypertension, stroke, or cancer, involve complex interactions between various systems/organs of the body and mutual influences, thus implicating a multiorgan approach. The dual-flow IVTech Live-Box2 bioreactor is a recently developed inter-connected dynamic cell culture model able to mimic organ crosstalk, since cells belonging to different organs can be connected and grown under flow conditions in a more physiological environment. This study aims to setup for the first time a 2-way connected culture of human neuroblastoma cells, SH-SY5Y, and Human Coronary Artery Smooth Muscle Cells, HCASMC through a dual-flow IVTech Live-Box2 bioreactor, in order to represent a simplified model of nervous-cardiovascular systems crosstalk, possibly relevant for the above-mentioned diseases. The system was tested by treating the cells with 10nM angiotensin II (AngII) inducing PKCβII/HuR/VEGF pathway activation, since AngII and PKCβII/HuR/VEGF pathway are relevant in cardiovascular and neuroscience research. Three different conditions were applied: 1- HCASMC and SH-SY5Y separately seeded in petri dishes (static condition); 2- the two cell lines separately seeded under flow (dynamic condition); 3- the two lines, seeded in dynamic conditions, connected, each maintaining its own medium, with a membrane as interface for biohumoral changes between the two mediums, and then treated. We detected that only in condition 3 there was a synergic AngII-dependent VEGF production in SH-SY5Y cells coupled to an AngII-dependent PKCβII/HuR/VEGF pathway activation in HCASMC, consistent with the observed physiological response in vivo. HCASMC response to AngII seems therefore to be generated by/derived from the reciprocal cell crosstalk under the dynamic inter-connection ensured by the dual flow LiveBox 2 bioreactor. This system can represent a useful tool for studying the crosstalk between organs, helpful for instance in rehabilitation research or when investigating chronic diseases; further, it offers the advantageous opportunity of cultivating each cell line in its own medium, thus mimicking, at least in part, distinct tissue milieu. © 2020 Marchesi et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
AB - Chronic conditions requiring long-term rehabilitation therapies, such as hypertension, stroke, or cancer, involve complex interactions between various systems/organs of the body and mutual influences, thus implicating a multiorgan approach. The dual-flow IVTech Live-Box2 bioreactor is a recently developed inter-connected dynamic cell culture model able to mimic organ crosstalk, since cells belonging to different organs can be connected and grown under flow conditions in a more physiological environment. This study aims to setup for the first time a 2-way connected culture of human neuroblastoma cells, SH-SY5Y, and Human Coronary Artery Smooth Muscle Cells, HCASMC through a dual-flow IVTech Live-Box2 bioreactor, in order to represent a simplified model of nervous-cardiovascular systems crosstalk, possibly relevant for the above-mentioned diseases. The system was tested by treating the cells with 10nM angiotensin II (AngII) inducing PKCβII/HuR/VEGF pathway activation, since AngII and PKCβII/HuR/VEGF pathway are relevant in cardiovascular and neuroscience research. Three different conditions were applied: 1- HCASMC and SH-SY5Y separately seeded in petri dishes (static condition); 2- the two cell lines separately seeded under flow (dynamic condition); 3- the two lines, seeded in dynamic conditions, connected, each maintaining its own medium, with a membrane as interface for biohumoral changes between the two mediums, and then treated. We detected that only in condition 3 there was a synergic AngII-dependent VEGF production in SH-SY5Y cells coupled to an AngII-dependent PKCβII/HuR/VEGF pathway activation in HCASMC, consistent with the observed physiological response in vivo. HCASMC response to AngII seems therefore to be generated by/derived from the reciprocal cell crosstalk under the dynamic inter-connection ensured by the dual flow LiveBox 2 bioreactor. This system can represent a useful tool for studying the crosstalk between organs, helpful for instance in rehabilitation research or when investigating chronic diseases; further, it offers the advantageous opportunity of cultivating each cell line in its own medium, thus mimicking, at least in part, distinct tissue milieu. © 2020 Marchesi et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
KW - ELAV like protein 1
KW - protein kinase C beta
KW - protein kinase C beta II
KW - unclassified drug
KW - vasculotropin
KW - Article
KW - cardiovascular system
KW - CASMC cell line
KW - cell culture
KW - cell density
KW - cell function
KW - cell separation
KW - cell viability
KW - controlled study
KW - enzyme linked immunosorbent assay
KW - human
KW - human cell
KW - in vivo study
KW - molecular dynamics
KW - MTT assay
KW - nervous system
KW - neuroscience
KW - SH-SY5Y cell line
KW - signal transduction
KW - Western blotting
KW - biological model
KW - bioreactor
KW - cell communication
KW - cytology
KW - metabolism
KW - nerve cell
KW - smooth muscle cell
KW - tumor cell line
KW - Bioreactors
KW - Cell Communication
KW - Cell Line, Tumor
KW - Humans
KW - Models, Cardiovascular
KW - Models, Neurological
KW - Myocytes, Smooth Muscle
KW - Neurons
KW - Signal Transduction
U2 - 10.1371/journal.pone.0242627
DO - 10.1371/journal.pone.0242627
M3 - Article
VL - 15
JO - PLoS ONE
JF - PLoS ONE
SN - 1932-6203
IS - 11 November
ER -