Ribosomal Protein L13a as a Reference Gene for Human Bone Marrow-Derived Mesenchymal Stromal Cells During Expansion, Adipo-, Chondro-, and Osteogenesis

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Author(s) Studer, Deborah, Lischer, Stefanie, Jochum, Wolfram, Ehrbar, Martin, Zenobi-Wong, Marcy, Maniura-Weber, Katharina
Publication Type Journal Items, Publication Status: Published
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Title Ribosomal Protein L13a as a Reference Gene for Human Bone Marrow-Derived Mesenchymal Stromal Cells During Expansion, Adipo-, Chondro-, and Osteogenesis
Author(s) Studer, Deborah
Lischer, Stefanie
Jochum, Wolfram
Ehrbar, Martin
Zenobi-Wong, Marcy
Maniura-Weber, Katharina
Journal or Series Title Tissue Engineering Part C-Methods
Volume Number 18
Issue Number 10
Start Page 761
End Page 771
ISSN 1937-3384
1937-3392
Publisher Mary Ann Liebert
Publication Place New Rochelle, NY
Publication Date 2012-10
Abstract In the field of human mesenchymal stromal cell (MSC) research, quantitative real-time reverse transcription-polymerase chain reaction (qPCR) is the method of choice to study changes in gene expression patterns upon differentiation, application of stimuli, or of factors such as inhibitors or siRNAs. To reliably detect small changes, the use of a reference gene (RG) that is stably expressed under all conditions is essential. The large number of different RGs used in the field and the lack of validation of their suitability make the comparison between studies impossible. Therefore, this work aims to establish one single RG for mesodermal differentiation studies that use MSCs. Seven commonly used RGs (glyceraldehyde-3-phosphate dehydrogenase [GAPDH], ribosomal protein L13a [RPL13a], beta-actin [ACTB], tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein, zeta-polypeptide [YWHAZ], eukaryotic translational elongation factor 1 alpha [EF1 alpha], beta 2-microglobulin [B2M], and 18S ribosomal RNA [18S]) were investigated concerning their mRNA expression stability during expansion of bone marrow-derived MSCs up to four passages as well as during their adipo-, chondro-, and osteogenenic differentiation on days 9, 16, and 22 after induction. RPL13a was validated for qPCR studies of MSCs (bone marrow-and placenta-derived) and, additionally, for primary human bone cells (HBCs) and the osteosarcoma cell line MG-63. GAPDH and ACTB, the two most frequently used RGs, showed the highest expression variance. The superior performance of RPL13a should make it the RG of choice for all MSC studies addressing mesodermal differentiation.
DOI 10.1089/ten.tec.2012.0081
Document Type Article
Publication Status Published
Language English
Assigned Organisational Unit(s) 03949
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NEBIS System Number 005828323
Source Database ID WOS-000309558600004
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@article{Stdr2012,
  author = "Studer, Deborah and Lischer, Stefanie and Jochum, Wolfram and Ehrbar, Martin and Zenobi-Wong, Marcy and Maniura-Weber, Katharina",
  title = "{R}ibosomal {P}rotein {L}13a as a {R}eference {G}ene for {H}uman {B}one {M}arrow-{D}erived {M}esenchymal {S}tromal {C}ells {D}uring {E}xpansion, {A}dipo-, {C}hondro-, and {O}steogenesis",
  journal = "Tissue Engineering Part C-Methods",
  year = 2012,
  volume = "18",
  number = "10",
  pages = "761--771",
  month = oct,
}


E-Citations record created: Mon, 12 Nov 2012, 06:55:13 CET