PMID- 24080205
OWN - NLM
STAT- MEDLINE
DCOM- 20140607
LR  - 20191210
IS  - 1876-7753 (Electronic)
IS  - 1873-5061 (Linking)
VI  - 11
IP  - 3
DP  - 2013 Nov
TI  - Identification of suitable reference genes in bone marrow stromal cells from 
      osteoarthritic donors.
PG  - 1288-98
LID - S1873-5061(13)00126-8 [pii]
LID - 10.1016/j.scr.2013.08.015 [doi]
AB  - Bone marrow stromal cells (BMSCs) are key cellular components for musculoskeletal 
      tissue engineering strategies. Furthermore, recent data suggest that BMSCs are 
      involved in the development of Osteoarthritis (OA) being a frequently occurring 
      degenerative joint disease. Reliable reference genes for the molecular evaluation 
      of BMSCs derived from donors exhibiting OA as a primary co-morbidity have not 
      been reported on yet. Hence, the aim of the study was to identify reference genes 
      suitable for comparative gene expression analyses using OA-BMSCs. Passage 1 bone 
      marrow derived BMSCs were isolated from n=13 patients with advanced stage 
      idiopathic hip osteoarthritis and n=15 age-matched healthy donors. The expression 
      of 31 putative reference genes was analyzed by quantitative reverse transcription 
      polymerase chain reaction (qRT-PCR) using a commercially available TaqMan((R)) 
      assay. Calculating the coefficient of variation (CV), mRNA expression stability 
      was determined and afterwards validated using geNorm and NormFinder algorithms. 
      Importin 8 (IPO8), TATA box binding protein (TBP), and cancer susceptibility 
      candidate 3 (CASC3) were identified as the most stable reference genes. Notably, 
      commonly used reference genes, e.g. beta-actin (ACTB) and beta-2-microglobulin 
      (B2M) were among the most unstable genes. For normalization of gene expression 
      data of OA-BMSCs the combined use of IPO8, TBP, and CASC3 gene is recommended.
CI  - (c) 2013.
FAU - Schildberg, Theresa
AU  - Schildberg T
AD  - University Centre for Orthopaedics & Trauma Surgery and Centre for Translational 
      Bone, Joint & Soft Tissue Research, University Hospital Carl Gustav Carus at the 
      Technische Universitat Dresden, Dresden, Germany. Electronic address: 
      theresa.schildberg@uniklinikum-dresden.de.
FAU - Rauh, Juliane
AU  - Rauh J
FAU - Bretschneider, Henriette
AU  - Bretschneider H
FAU - Stiehler, Maik
AU  - Stiehler M
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20130909
PL  - England
TA  - Stem Cell Res
JT  - Stem cell research
JID - 101316957
RN  - 0 (CASC3 protein, human)
RN  - 0 (IPO8 protein, human)
RN  - 0 (Neoplasm Proteins)
RN  - 0 (Nuclear Proteins)
RN  - 0 (RNA, Messenger)
RN  - 0 (RNA-Binding Proteins)
RN  - 0 (TATA-Box Binding Protein)
RN  - 0 (beta Karyopherins)
SB  - IM
MH  - Aged
MH  - Algorithms
MH  - Bone Marrow Cells/*cytology
MH  - Gene Expression
MH  - Humans
MH  - Mesenchymal Stem Cells/cytology/*metabolism
MH  - Middle Aged
MH  - Neoplasm Proteins/*genetics/metabolism
MH  - Nuclear Proteins/*genetics/metabolism
MH  - Osteoarthritis/*genetics/metabolism/*pathology
MH  - RNA, Messenger/metabolism
MH  - RNA-Binding Proteins
MH  - Real-Time Polymerase Chain Reaction
MH  - TATA-Box Binding Protein/*genetics/metabolism
MH  - beta Karyopherins/*genetics/metabolism
EDAT- 2013/10/02 06:00
MHDA- 2014/06/08 06:00
CRDT- 2013/10/02 06:00
PHST- 2013/03/27 00:00 [received]
PHST- 2013/08/20 00:00 [revised]
PHST- 2013/08/31 00:00 [accepted]
PHST- 2013/10/02 06:00 [entrez]
PHST- 2013/10/02 06:00 [pubmed]
PHST- 2014/06/08 06:00 [medline]
AID - S1873-5061(13)00126-8 [pii]
AID - 10.1016/j.scr.2013.08.015 [doi]
PST - ppublish
SO  - Stem Cell Res. 2013 Nov;11(3):1288-98. doi: 10.1016/j.scr.2013.08.015. Epub 2013 
      Sep 9.