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Stable expression of a neuronal dopaminergic progenitor phenotype in cell lines derived from human amniotic fluid cells
Author(s) -
McLaughlin David,
Tsirimonaki Emmanouella,
Vallianatos George,
Sakellaridis Nikolaos,
Chatzistamatiou Theofanis,
StavropoulosGioka Catherine,
Tsezou Aspasia,
Messinis Ioannis,
Mangoura Dimitra
Publication year - 2006
Publication title -
journal of neuroscience research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.72
H-Index - 160
eISSN - 1097-4547
pISSN - 0360-4012
DOI - 10.1002/jnr.20828
Subject(s) - biology , dopaminergic , progenitor cell , microbiology and biotechnology , embryonic stem cell , population , nestin , stem cell , neural stem cell , dopamine , neuroscience , genetics , medicine , environmental health , gene
Abstract Cells from human amniotic fluid derived from the fetus are considered a source of multipotent cells. Their properties have not been fully exploited, partially because unlike other embryonic sources such as embryonic stem (ES) cells, cell lines from amniocentesis samples have not been generated. We have established and characterized the properties of eight individual cell lines. Flow cytometry using several cell surface markers showed that all cell lines generated consisted of homogeneous populations that lack HLAII antigenicity. Using a combination of immunocytochemistry, Western blotting, and RT‐PCR, we found weak expression of Oct4 and nestin and strong expression of tubulin‐βIII, MAP2, and tau. Specific markers for cholinergic, (nor)adrenergic, and GABAergic neurons or glia were weakly expressed or absent, whereas expression of factors implicated in early induction of dopaminergic neurons, TGF‐β3 and β‐catenin were present. Further analysis showed strong expression of EN‐1, c‐RET, PTX3, and NURR1 essential for induction and survival of midbrain dopaminergic neurons, TH, AADC, and VMAT2 components of dopamine synthesis and secretion, and syntaxin1A and SNAP‐25 necessary for neurotransmitter exocytosis. This phenotype was retained throughout passages and up to the current passage 36. Expression of neuronal and dopaminergic markers in individual AF cell lines was comparable to expression in neurons induced from ES cells and in IMR‐32 and SH‐SY5Y neuroblastomas. Our data show that cell lines can be derived from subcultures of amniocentesis, and are primarily composed of a population of progenitors with a phenotype similar to that of committed mesencephalic dopaminergic neurons. © 2006 Wiley‐Liss, Inc.