Open AccessCell Renewing in Neuroblastoma: Electrophysiological and Immunocytochemical Characterization of Stem Cells and Derivatives
Author(s) -
Biagiotti Tiziana,
D'Amico Massimo,
Marzi Ilaria,
Di Gennaro Paola,
Arcangeli Annarosa,
Wanke Enzo,
Olivotto Massimo
Publication year - 2006
Publication title -
stem cells
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.159
H-Index - 229
eISSN - 1549-4918
pISSN - 1066-5099
DOI - 10.1634/stemcells.2004-0264
Subject(s) - biology , stem cell , neural crest , microbiology and biotechnology , clone (java method) , electrophysiology , clonogenic assay , cellular differentiation , sox2 , population , calponin , cell culture , neuroscience , genetics , actin , transcription factor , embryo , dna , demography , sociology , gene
We explored the stem cell compartment of the SH‐SY5Y neuroblastoma (NB) clone and its development by a novel approach, integrating clonal and immunocytochemical investigations with patch‐clamp measurements of ion currents simultaneously expressed on single cells. The currents selected were the triad I HERG , I KDR , I Na , normally expressed at varying mutual ratios during development of neural crest stem cells, from which NB derives upon neoplastic transformation. These ratios could be used as electrophysiological clusters of differentiation (ECDs), identifying otherwise indistinguishable stages in maturation. Subcloning procedures allowed the isolation of highly clonogenic substrate‐adherent (S‐type) cells that proved to be p75‐ and nestinpositive and were characterized by a nude electrophysiological profile (ECDS 0 ). These cells expressed negligible levels of the triad and manifested the capacity of generating the two following lineages: first, a terminally differentiating, smooth muscular lineage, positive for calponin and smooth muscle actin, whose electrophysiological profile is characterized by a progressive diminution of I HERG , the increase of I KDR and I Na , and the acquisition of I KIR (ECDS 2 ); second, a neuronal abortive pathway (NF‐68 positive), characterized by a variable expression of I HERG and I KDR and a low expression of I Na (ECDN S ). This population manifested a vigorous amplification, monopolizing the stem cell compartment at the expense of the smooth muscular lineage to such an extent that neuronal‐like (N‐type) cells must be continuously removed if the latter are to develop.