2-DE proteome analysis of a proliferating and differentiating human neuronal stem cell line (ReNcell VM).
The proteome of a proliferating human stem cell line was analyzed and then utilized to detect stem cell differentiation-associated changes in the protein profile. The analysis was conducted with a stable human fetal midbrain stem cell line (ReNcell VM) that displays the properties of a neural stem cell. Therefore, acquisition of proteomic data should be representative of cultured human neural stem cells (hNSCs) in general. Here we present a 2-DE protein-map of this cell line with annotations of 402 spots representing 318 unique proteins identified by MS. The subsequent proteome profiling of differentiating cells of this stem cell line at days 0, 4 and 7 of differentiation revealed changes in the expression of 49 identified spots that could be annotated to 45 distinct proteins. This differentiation-associated expression pattern was validated by Western blot analysis for transgelin-2, proliferating cell nuclear antigen, as well as peroxiredoxin 1 and 4. The group of regulated proteins also included NudC, ubiquilin-1, STRAP, stress-70 protein, creatine kinase B, glial fibrillary acidic protein and vimentin. Our results reflect the large rearrangement of the proteome during the differentiation process of the stem cells to terminally differentiated neurons and offer the possibility for further characterization of specific targets driving the stem cell differentiation.
Hoffrogge
Raimund
R
Neurobiological Laboratory, Department of Neurology, Medical Faculty, University of Rostock, Germany.
Mikkat
Stefan
S
Scharf
Christian
C
Beyer
Susanne
S
Christoph
Hilmar
H
Pahnke
Jens
J
Mix
Eilhard
E
Berth
Matthias
M
Uhrmacher
Adelinde
A
Zubrzycki
Igor Z
IZ
Miljan
Erik
E
Völker
Uwe
U
Rolfs
Arndt
A
D015153:Blotting, Western; D002454:Cell Differentiation; D002460:Cell Line; D002461:Cell Line, Transformed; D049109:Cell Proliferation; D002472:Cell Transformation, Viral; D019295:Computational Biology; D003470:Culture Media / Q000737:chemistry / Q000494:pharmacology; D030562:Databases, Protein; D015180:Electrophoresis, Gel, Two-Dimensional; D004815:Epidermal Growth Factor / Q000494:pharmacology; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D005819:Genetic Markers; D006801:Humans; D013058:Mass Spectrometry; D008636:Mesencephalon / Q000166:cytology / Q000196:embryology; D008840:Microfilament Proteins / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism; D009124:Muscle Proteins / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism; D009363:Neoplasm Proteins / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism; D009474:Neurons / Q000166:cytology*; D010449:Peptide Mapping; D010544:Peroxidases / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism; D054464:Peroxiredoxins; D018809:Proliferating Cell Nuclear Antigen / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism; D020543:Proteome / Q000032:analysis*; D012190:Retroviridae / Q000235:genetics; D012641:Selection, Genetic; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013234:Stem Cells / Q000166:cytology / Q000502:physiology*; D014161:Transduction, Genetic; D019076:Transgenes; D014357:Trypsin / Q000494:pharmacology
D003470:Culture Media; D005819:Genetic Markers; D008840:Microfilament Proteins; D009124:Muscle Proteins; D009363:Neoplasm Proteins; D018809:Proliferating Cell Nuclear Antigen; D020543:Proteome; C051615:transgelin; D016222:Fibroblast Growth Factor 2; D004815:Epidermal Growth Factor; D010544:Peroxidases; C515940:PRDX1 protein, human; C515980:PRDX4 protein, human; D054464:Peroxiredoxins; D014357:Trypsin