SH-SY5Y is a human derived cell line used in scientific research. The original cell line, called SK-N-SH, from which it was subcloned was isolated from a bone marrow biopsy taken from a four-year-old female with neuroblastoma. SH-SY5Y cells are often used as in vitro models of neuronal function and differentiation. They are adrenergic in phenotype but also express dopaminergic markers and, as such, have been used to study Parkinson's disease, neurogenesis, and other characteristics of brain cells.
SH-SY5Y cells can form mounds of undifferentiated cells, which then spread differentiated cells into the surrounding area. This growth formation can be referred to as the "over-easy formation"
SH-SY5Y was cloned from a bone marrow biopsy derived line called SK-N-SH by the laboratory of June Biedler and first reported in 1973.[1] A neuroblast-like subclone of SK-N-SH, named SH-SY, was subcloned as SH-SY5, which was subcloned a third time to produce the SH-SY5Y line, first described in 1978.[2] The cloning process involved the selection of individual cells or clusters expressing neuron-like characteristics. The SH-SY5Y line is genetically female with two X and no Y chromosome, as expected given the origin from a four-year-old female.
Morphologyedit
The cells typically grow in tissue culture in two distinct ways. Some grow into clumps of cells which float in the media, while others form clumps which stick to the dish. SH-SY5Y cells can spontaneously interconvert between two phenotypes in vitro, the neuroblast-like cells and the Epithelial like cells, although the mechanisms underlying this process are not understood.[3] However, the cell line is appreciated to be N-type (neuronal), given its morphology and the ability to differentiate the cells into along the neuronal lineage (in contrast to the S-type SH-EP subcloned cell line, also derived from SK-N-SH).[4] Cells with short spiny neurite-like processes migrate out from these adherent clumps. SH-SY5Y cells possess an abnormal chromosome 1, where there is an additional copy of a 1q segment and is referred to trisomy 1q. SH-SY5Y cells are known to be dopamine beta hydroxylase active, acetylcholinergic, glutamatergic and adenosinergic. The cells have very different growth phases, outlined in the surrounding pictures. The cells both propagate via mitosis and differentiate by extending neurites to the surrounding area. While dividing, the aggregated cells can look so different from the differentiated cells following the K'annul-index tumor cellularity (TNF, for Tumor Necrosis Factor), that new scientists often mistake one or the other for contamination. The dividing cells can form clusters of cells which are reminders of their cancerous nature, but certain treatments such as retinoic acid, BDNF, or TPA can force the cells to dendrify and differentiate. Moreover, induction by retinoic acid results in inhibition of cell growth and enhanced production of noradrenaline from SH-SY5Y cells [5][6]
The SH-SY5Y cells are dense and considered to be confluent at this stage. Neuritic growth is more pronounced on the lower part of the photo
Media and Cultivationedit
The most common growing cocktail used is a 1:1 mixture of DMEM and Ham's F12 medium and 10% supplemental fetal bovine serum. The DMEM usually contains 3.7 g/L sodium bicarbonate, 2 mM L-Glutamine, 1 mM sodium pyruvate and 0.1 mM nonessential amino acids.[7] The cells are always grown at 37 degrees Celsius with 95% air and 5% carbon dioxide. It is advised to cultivate the cells in flasks which are coated for cell culture adhesion, this will aid in differentiation and dendrification of the neuroblastoma. In general the cells are quite robust and will grow in most widely used tissue culture media. However, it has been recently established that DMEM is superior to DMEM:F12 for propagation of SH-SY5Y.[8]
The doubling time of SH-SY5Y cells during the exponential phase is 67.3 h ± 5.8 h. The average generation time (G) was 4.23 ± 0.84 d, the division rate per cell unit (r) was 0.25 ± 0.05, and the multiplication factor (V) was 3.49 ± 1.21. SH-SY5Y cells show growth-related parameters that were typical or even superior to many described immortalized tumorderived cell lines. The proliferation capacity is stable over time and also beyond passage 10.[9]
SH-SY5Y has a dopamine-β-hydroxylase activity and can convert dopamine to norepinephrine. It will also form tumors in nude mice in around 3–4 weeks. The loss of neuronal characteristics has been described with increasing passage numbers. Therefore, it is recommended not to be used after passage 20 or verify specific characteristics such as noradrenaline uptake or neuronal tumor markers.
Referencesedit
^Biedler JL, Helson L, Spengler BA (November 1973). "Morphology and growth, tumorigenicity, and cytogenetics of human neuroblastoma cells in continuous culture". Cancer Res. 33 (11): 2643–52. PMID 4748425.
^Biedler JL, Roffler-Tarlov S, Schachner M, Freedman LS (November 1978). "Multiple neurotransmitter synthesis by human neuroblastoma cell lines and clones". Cancer Res. 38 (11 Pt 1): 3751–7. PMID 29704.
^Feles, Sebastian; Overath, Christian; Reichardt, Sina; Diegeler, Sebastian; Schmitz, Claudia; Kronenberg, Jessica; Baumstark-Khan, Christa; Hemmersbach, Ruth; Hellweg, Christine E.; Liemersdorf, Christian (August 2022). "Streamlining Culture Conditions for the Neuroblastoma Cell Line SH-SY5Y: A Prerequisite for Functional Studies". Methods and Protocols. 5 (4): 58. doi:10.3390/mps5040058. PMC9326679. PMID 35893584.
^La Quaglia Michael P.; Manchester Karen M. (1996). "A comparative analysis of neuroblastic and substrate-adherent human neuroblastoma cell lines". Journal of Pediatric Surgery. 31 (2): 315–318. doi:10.1016/S0022-3468(96)90025-1. PMID 8938368.
^Kunzler A, Zeidán-Chuliá F, Gasparotto J, Girardi CS, Klafke K, Petiz LL, Bortolin RC, Rostirolla DC, Zanotto-Filho A, de Bittencourt Pasquali MA, Dickson P, Dunkley P, Moreira JC, Gelain DP (2017). "Changes in Cell Cycle and Up-Regulation of Neuronal Markers During SH-SY5Y Neurodifferentiation by Retinoic Acid are Mediated by Reactive Species Production and Oxidative Stress". Mol. Neurobiol. 54 (9): 6903–6916. doi:10.1007/s12035-016-0189-4. PMID 27771902. S2CID 3515130.
^Feles, Sebastian; Overath, Christian; Reichardt, Sina; Diegeler, Sebastian; Schmitz, Claudia; Kronenberg, Jessica; Baumstark-Khan, Christa; Hemmersbach, Ruth; Hellweg, Christine E.; Liemersdorf, Christian (12 July 2022). "Streamlining Culture Conditions for the Neuroblastoma Cell Line SH-SY5Y: A Prerequisite for Functional Studies". Methods and Protocols. 5 (4): 58. doi:10.3390/mps5040058. PMC9326679. PMID 35893584.
^Sakagami, Hiroshi; Suzuki, Ryuichiro; Shirataki, Yoshiaki; Iwama, Soichi; Nakagawa, Mika; Suzuki, Hayato; Tanaka, Kenta; Tamura, Nobuaki; Takeshima, Hiroshi (3 November 2017). "Re-evaluation of Culture Condition of PC12 and SH-SY5Y Cells Based on Growth Rate and Amino Acid Consumption". In Vivo. 31 (6): 1089–1095. doi:10.21873/invivo.11174. PMC5756636. PMID 29102930.
^Feles, Sebastian; Overath, Christian; Reichardt, Sina; Diegeler, Sebastian; Schmitz, Claudia; Kronenberg, Jessica; Baumstark-Khan, Christa; Hemmersbach, Ruth; Hellweg, Christine E.; Liemersdorf, Christian (12 July 2022). "Streamlining Culture Conditions for the Neuroblastoma Cell Line SH-SY5Y: A Prerequisite for Functional Studies". Methods and Protocols. 5 (4): 58. doi:10.3390/mps5040058. PMC9326679. PMID 35893584.
ATCC- American Type Culture Collection, a source of SH-SY5Y cells and other biologicials
ATTC product data for SH-SY5Y cells, catalog CRL-2266
Sigma Product Page
Canals, Meritxell; Angulo, Ester; Casado, Vicent; Canela, Enric I.; Mallol, Josefa; Vinals, Francesc; Staines, William; Tinner, Barbro; Agnati, Luigi; Fuxe, Kjell; Ferre, Sergi; Lluis, Carmen; Franco, Rafael; Franco, R (January 2005). "Molecular mechanisms involved in the adenosine A1 and A2A receptor-induced neuronal differentiation in neuroblastoma cells and striatal primary cultures". Journal of Neurochemistry. 92 (2): 337–348. doi:10.1111/j.1471-4159.2004.02856.x. PMID 15663481. S2CID 2766681.
Hillion, Joëlle; Canals, Meritxell; Torvinen, Maria; Casadó, Vicent; Scott, Rizaldy; Terasmaa, Anton; Hansson, Anita; Watson, Stanley; Olah, Mark E.; Mallol, Josefa; Canela, Enric I.; Zoli, Michele; Agnati, Luigi F.; Ibáñez, Carlos F.; Lluis, Carme; Franco, Rafael; Ferré, Sergi; Fuxe, Kjell (17 May 2002). "Coaggregation, Cointernalization, and Codesensitization of Adenosine A 2A Receptors and Dopamine D 2 Receptors". Journal of Biological Chemistry. 277 (20): 18091–18097. doi:10.1074/jbc.M107731200. PMID 11872740.
Okuda, Katsuhiro; Kotake, Yaichiro; Ohta, Shigeru (2006). "Parkinsonism-Preventing Activity of 1-Methyl-1,2,3,4-tetrahydroisoquinoline Derivatives in C57BL Mouse in Vivo". Biological & Pharmaceutical Bulletin. 29 (7): 1401–1403. doi:10.1248/bpb.29.1401. PMID 16819177.
December 15, 2023
sy5y, human, derived, cell, line, used, scientific, research, original, cell, line, called, from, which, subcloned, isolated, from, bone, marrow, biopsy, taken, from, four, year, female, with, neuroblastoma, cells, often, used, vitro, models, neuronal, functio. SH SY5Y is a human derived cell line used in scientific research The original cell line called SK N SH from which it was subcloned was isolated from a bone marrow biopsy taken from a four year old female with neuroblastoma SH SY5Y cells are often used as in vitro models of neuronal function and differentiation They are adrenergic in phenotype but also express dopaminergic markers and as such have been used to study Parkinson s disease neurogenesis and other characteristics of brain cells SH SY5Y cells can form mounds of undifferentiated cells which then spread differentiated cells into the surrounding area This growth formation can be referred to as the over easy formation Contents 1 History 2 Morphology 3 Media and Cultivation 4 References 5 External linksHistory editSH SY5Y was cloned from a bone marrow biopsy derived line called SK N SH by the laboratory of June Biedler and first reported in 1973 1 A neuroblast like subclone of SK N SH named SH SY was subcloned as SH SY5 which was subcloned a third time to produce the SH SY5Y line first described in 1978 2 The cloning process involved the selection of individual cells or clusters expressing neuron like characteristics The SH SY5Y line is genetically female with two X and no Y chromosome as expected given the origin from a four year old female Morphology editThe cells typically grow in tissue culture in two distinct ways Some grow into clumps of cells which float in the media while others form clumps which stick to the dish SH SY5Y cells can spontaneously interconvert between two phenotypes in vitro the neuroblast like cells and the Epithelial like cells although the mechanisms underlying this process are not understood 3 However the cell line is appreciated to be N type neuronal given its morphology and the ability to differentiate the cells into along the neuronal lineage in contrast to the S type SH EP subcloned cell line also derived from SK N SH 4 Cells with short spiny neurite like processes migrate out from these adherent clumps SH SY5Y cells possess an abnormal chromosome 1 where there is an additional copy of a 1q segment and is referred to trisomy 1q SH SY5Y cells are known to be dopamine beta hydroxylase active acetylcholinergic glutamatergic and adenosinergic The cells have very different growth phases outlined in the surrounding pictures The cells both propagate via mitosis and differentiate by extending neurites to the surrounding area While dividing the aggregated cells can look so different from the differentiated cells following the K annul index tumor cellularity TNF for Tumor Necrosis Factor that new scientists often mistake one or the other for contamination The dividing cells can form clusters of cells which are reminders of their cancerous nature but certain treatments such as retinoic acid BDNF or TPA can force the cells to dendrify and differentiate Moreover induction by retinoic acid results in inhibition of cell growth and enhanced production of noradrenaline from SH SY5Y cells 5 6 nbsp The SH SY5Y cells are dense and considered to be confluent at this stage Neuritic growth is more pronounced on the lower part of the photoMedia and Cultivation editThe most common growing cocktail used is a 1 1 mixture of DMEM and Ham s F12 medium and 10 supplemental fetal bovine serum The DMEM usually contains 3 7 g L sodium bicarbonate 2 mM L Glutamine 1 mM sodium pyruvate and 0 1 mM nonessential amino acids 7 The cells are always grown at 37 degrees Celsius with 95 air and 5 carbon dioxide It is advised to cultivate the cells in flasks which are coated for cell culture adhesion this will aid in differentiation and dendrification of the neuroblastoma In general the cells are quite robust and will grow in most widely used tissue culture media However it has been recently established that DMEM is superior to DMEM F12 for propagation of SH SY5Y 8 The doubling time of SH SY5Y cells during the exponential phase is 67 3 h 5 8 h The average generation time G was 4 23 0 84 d the division rate per cell unit r was 0 25 0 05 and the multiplication factor V was 3 49 1 21 SH SY5Y cells show growth related parameters that were typical or even superior to many described immortalized tumorderived cell lines The proliferation capacity is stable over time and also beyond passage 10 9 SH SY5Y has a dopamine b hydroxylase activity and can convert dopamine to norepinephrine It will also form tumors in nude mice in around 3 4 weeks The loss of neuronal characteristics has been described with increasing passage numbers Therefore it is recommended not to be used after passage 20 or verify specific characteristics such as noradrenaline uptake or neuronal tumor markers References edit Biedler JL Helson L Spengler BA November 1973 Morphology and growth tumorigenicity and cytogenetics of human neuroblastoma cells in continuous culture Cancer Res 33 11 2643 52 PMID 4748425 Biedler JL Roffler Tarlov S Schachner M Freedman LS November 1978 Multiple neurotransmitter synthesis by human neuroblastoma cell lines and clones Cancer Res 38 11 Pt 1 3751 7 PMID 29704 Feles Sebastian Overath Christian Reichardt Sina Diegeler Sebastian Schmitz Claudia Kronenberg Jessica Baumstark Khan Christa Hemmersbach Ruth Hellweg Christine E Liemersdorf Christian August 2022 Streamlining Culture Conditions for the Neuroblastoma Cell Line SH SY5Y A Prerequisite for Functional Studies Methods and Protocols 5 4 58 doi 10 3390 mps5040058 PMC 9326679 PMID 35893584 La Quaglia Michael P Manchester Karen M 1996 A comparative analysis of neuroblastic and substrate adherent human neuroblastoma cell lines Journal of Pediatric Surgery 31 2 315 318 doi 10 1016 S0022 3468 96 90025 1 PMID 8938368 Girardi CS Rostirolla DC Lini FJ Brum PO Delgado J Ribeiro CT Teixeira AA Peixoto DO Heimfarth L Kunzler A Moreira JC Gelain DP 2019 Nuclear RXRa and RXRb receptors exert distinct and opposite effects on RA mediated neuroblastoma differentiation Biochimica et Biophysica Acta BBA Molecular Cell Research 1866 3 317 328 doi 10 1016 j bbamcr 2018 11 014 PMID 30529222 Kunzler A Zeidan Chulia F Gasparotto J Girardi CS Klafke K Petiz LL Bortolin RC Rostirolla DC Zanotto Filho A de Bittencourt Pasquali MA Dickson P Dunkley P Moreira JC Gelain DP 2017 Changes in Cell Cycle and Up Regulation of Neuronal Markers During SH SY5Y Neurodifferentiation by Retinoic Acid are Mediated by Reactive Species Production and Oxidative Stress Mol Neurobiol 54 9 6903 6916 doi 10 1007 s12035 016 0189 4 PMID 27771902 S2CID 3515130 Feles Sebastian Overath Christian Reichardt Sina Diegeler Sebastian Schmitz Claudia Kronenberg Jessica Baumstark Khan Christa Hemmersbach Ruth Hellweg Christine E Liemersdorf Christian 12 July 2022 Streamlining Culture Conditions for the Neuroblastoma Cell Line SH SY5Y A Prerequisite for Functional Studies Methods and Protocols 5 4 58 doi 10 3390 mps5040058 PMC 9326679 PMID 35893584 Sakagami Hiroshi Suzuki Ryuichiro Shirataki Yoshiaki Iwama Soichi Nakagawa Mika Suzuki Hayato Tanaka Kenta Tamura Nobuaki Takeshima Hiroshi 3 November 2017 Re evaluation of Culture Condition of PC12 and SH SY5Y Cells Based on Growth Rate and Amino Acid Consumption In Vivo 31 6 1089 1095 doi 10 21873 invivo 11174 PMC 5756636 PMID 29102930 Feles Sebastian Overath Christian Reichardt Sina Diegeler Sebastian Schmitz Claudia Kronenberg Jessica Baumstark Khan Christa Hemmersbach Ruth Hellweg Christine E Liemersdorf Christian 12 July 2022 Streamlining Culture Conditions for the Neuroblastoma Cell Line SH SY5Y A Prerequisite for Functional Studies Methods and Protocols 5 4 58 doi 10 3390 mps5040058 PMC 9326679 PMID 35893584 External links editThis article includes a list of general references but it lacks sufficient corresponding inline citations Please help to improve this article by introducing more precise citations October 2020 Learn how and when to remove this template message Cellosaurus entry for SH SY5Y ATCC American Type Culture Collection a source of SH SY5Y cells and other biologicials ATTC product data for SH SY5Y cells catalog CRL 2266 Sigma Product Page Canals Meritxell Angulo Ester Casado Vicent Canela Enric I Mallol Josefa Vinals Francesc Staines William Tinner Barbro Agnati Luigi Fuxe Kjell Ferre Sergi Lluis Carmen Franco Rafael Franco R January 2005 Molecular mechanisms involved in the adenosine A1 and A2A receptor induced neuronal differentiation in neuroblastoma cells and striatal primary cultures Journal of Neurochemistry 92 2 337 348 doi 10 1111 j 1471 4159 2004 02856 x PMID 15663481 S2CID 2766681 Hillion Joelle Canals Meritxell Torvinen Maria Casado Vicent Scott Rizaldy Terasmaa Anton Hansson Anita Watson Stanley Olah Mark E Mallol Josefa Canela Enric I Zoli Michele Agnati Luigi F Ibanez Carlos F Lluis Carme Franco Rafael Ferre Sergi Fuxe Kjell 17 May 2002 Coaggregation Cointernalization and Codesensitization of Adenosine A 2A Receptors and Dopamine D 2 Receptors Journal of Biological Chemistry 277 20 18091 18097 doi 10 1074 jbc M107731200 PMID 11872740 Okuda Katsuhiro Kotake Yaichiro Ohta Shigeru 2006 Parkinsonism Preventing Activity of 1 Methyl 1 2 3 4 tetrahydroisoquinoline Derivatives in C57BL Mouse in Vivo Biological amp Pharmaceutical Bulletin 29 7 1401 1403 doi 10 1248 bpb 29 1401 PMID 16819177 Retrieved from https en wikipedia org w index php title SH SY5Y amp oldid 1157450549, wikipedia, wiki, book, books, library,
