EXPRESSION OF NEURONAL CHARACTERISTICS IN HUMAN NEUROBLASTOMA CELL LINES, SK-N-BE(2) M17 AND SH-N-SH-SY5Y.
Item
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Title
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EXPRESSION OF NEURONAL CHARACTERISTICS IN HUMAN NEUROBLASTOMA CELL LINES, SK-N-BE(2) M17 AND SH-N-SH-SY5Y.
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Identifier
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AAI8302498
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identifier
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8302498
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Creator
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CARROLL, JOANNE MARLENE.
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Contributor
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Katherine Lyser
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Date
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1982
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Language
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English
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Publisher
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City University of New York.
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Subject
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Biology, General
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Abstract
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Two human neuroblastoma clonal cell lines, SK-N-BE(2) M17 and SK-N-SH-SY5Y, were passaged and maintained in serum-free hormone supplemented medium, N2 (Bottenstein & Sato, 1979). Both cell lines exhibited slower growth in N2 as compared to fetal calf serum (FCS) supplementation, however, the morphology, ultrastructure and qualitative expression of neurotransmitter synthesizing enzymes, tyrosine hydroxylase (TH) and dopamine-(beta)-hydroxylase (DBH), were similar.;Cultures of M17 and SY5Y in N2 medium expressed basal levels of TH lower than cultures in FCS medium. Addition of adenosine 3',5'-cyclic monophosphate (cyclic AMP) to N2 medium elevated the specific activity of TH to the level observed in serum medium. When added to FCS medium, cyclic AMP had no effect on TH activity. Growth of these cells in N2 medium revealed the presence of a cyclic AMP-dependent process in the regulation of TH.;Cyclic AMP also increased the outgrowth of long neurites in N2 but not in FCS medium. The ultrastructure of processes studied under all conditions, however, was similar.;Nerve growth factor (NGF) treatment of SY5Y cells increases the number of long neurites. This response requires serum factors and is not observed in N2 medium (Burmeister, 1982). Despite the morphological response, NGF at concentrations up to 100 ng/ml failed to affect the specific activity of TH or DBH in either SY5Y or M17.;While both cell lines expressed enzymes specific to noradrenergic cells, as determined by activity assay and immunocytochemical staining, their ultrastructure resembled early neuroblasts of the developing sympathetic nervous system. No mature cytoplasmic features, axonal or synaptic profiles were observed. Dense core vesicles were present, however, in both cell lines under all medium conditions studied. The content of the vesicles remains to be determined.;Defined medium N2 provided an adequate medium for maintenance of human neuroblastoma lines M17 and SY5Y. In addition, it revealed effects of cyclic AMP treatment on morphology and enzyme activity not observed in the presence of serum. This medium provides an improved system in which the role of factor(s) regulating specific neuronal functions may be assayed without complications introduced by serum.
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Type
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dissertation
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Source
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PQT Legacy CUNY.xlsx
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degree
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Ph.D.
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Program
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Biology
