Variations of protein levels in human amniotic fluid stem cells CD117/2 over passages 5-25.
Stability of cell lines is the prerequisite for all in vitro research, but literature on the stability of protein expression over passages is limited. Determination of specific stability markers, karyotyping, and morphology may not provide full information on this subject. It was the aim of the study to test protein level fluctuations in a human amniotic fluid stem cell line from passages 5, 7, 11, and 25. While karyotype, cell cycle, apoptosis rate, and 10 markers for characterization of the cell line remained unchanged (carried out at passages 5 and 25), cell volume was increased at passage 25. Significant protein fluctuations were observed for signaling, antioxidant, guidance cue, proteasomal, connective tissue, cytoskeleton proteins, chaperones, a chloride channel, and prothymosin at passages 5, 7, 11, and 25. Herein, the use of this gel-based proteomic screen, checking protein stability for the characterization of cell lines in addition to corresponding published markers, is proposed, in particular when experiments are run over several passages.
Department of Pediatrics, Medical University of Vienna, Wahringer Gurtel 18, 1090 Vienna, Austria.
Wei-Qiang
WQ
Chen
Nicol
N
Siegel
Lin
L
Li
Arnold
A
Pollak
Markus
M
Hengstschläger
Gert
G
Lubec
D000653:Amniotic Fluid / Q000166:cytology*; D015415:Biomarkers / Q000378:metabolism*; D018929:Cell Culture Techniques; D002460:Cell Line; D006801:Humans; D008969:Molecular Sequence Data; D019009:Proto-Oncogene Proteins c-kit / Q000378:metabolism*; D013234:Stem Cells / Q000737:chemistry* / Q000166:cytology / Q000502:physiology*
D015415:Biomarkers; D019009:Proto-Oncogene Proteins c-kit