PMID- 22698443
OWN - NLM
STAT- MEDLINE
DCOM- 20121227
LR  - 20161018
IS  - 1522-9653 (Electronic)
IS  - 1063-4584 (Linking)
VI  - 20
IP  - 9
DP  - 2012 Sep
TI  - Genomic chondrocyte culture profiling by array-CGH, interphase-FISH and RT-PCR.
PG  - 1039-45
LID - 10.1016/j.joca.2012.05.012 [doi]
AB  - OBJECTIVE: In vitro expansion is an important step to acquire sufficient cells in 
      human tissue engineering technologies. The high number of chondrocytes needed for 
      human articular cartilage implants requires in vitro expansion of the primary 
      cells, bearing a theoretical risk of in vitro induced changes in the genomes. To 
      gain more insights into this situation, model cultures were prepared and 
      analyzed. DESIGN: 25 chondrocyte cell DNA samples from nine donors were analyzed 
      by array comparative genomic hybridization (aCGH) on whole genome level and 28 
      chondrocyte cell samples from 16 individuals were analyzed by fluorescence in 
      situ hybridization (FISH) on single cell level. The expanded cells were further 
      characterized upon the chondrocytic mRNA phenotype by reverse-transciptase 
      polymerase chain reaction (RT-PCR). RESULTS: The molecular karyotyping results 
      revealed autosomal stability, but all male samples analyzed by aCGH displayed a 
      variable loss of the Y-chromosome. These data were confirmed by FISH-experiments 
      and suggest an age dependant effect toward the loss of the Y-chromosome in 
      cultured chondrocytes. RT-PCR data for the mRNAs from collagen types I, II, and 
      aggrecan and the pro-inflammatory cytokine interleukin-1ss (IL-1ss) did not reveal 
      any correlation of transcriptional activity in cultures with Y-chromosome losses, 
      nor were there statistically significant differences between cells from female 
      and male donors. CONCLUSIONS: While cells of male origin may suffer from an 
      age-related loss of the Y-chromosome, there was no indication of a functional 
      impairment. The data suggest some caution toward applying proliferative steps 
      when considering chondrocytes from elderly male patients for tissue engineering 
      approaches.
CI  - Copyright (c) 2012 Osteoarthritis Research Society International. Published by 
      Elsevier Ltd. All rights reserved.
FAU - Stumm, M
AU  - Stumm M
AD  - BG Berlin-Genetics GmbH, MDC-Buch, Berlin, Germany. stumm@kudamm-199.de
FAU - Boger, E
AU  - Boger E
FAU - Gaissmaier, C G
AU  - Gaissmaier CG
FAU - Osswald, C
AU  - Osswald C
FAU - Blankenburg, M
AU  - Blankenburg M
FAU - Wegner, R D
AU  - Wegner RD
FAU - Mollenhauer, J A
AU  - Mollenhauer JA
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20120612
PL  - England
TA  - Osteoarthritis Cartilage
JT  - Osteoarthritis and cartilage
JID - 9305697
RN  - 0 (Aggrecans)
RN  - 0 (Collagen Type I)
RN  - 0 (Collagen Type II)
RN  - 0 (Interleukin-1beta)
RN  - 0 (RNA, Messenger)
SB  - IM
MH  - Adult
MH  - Aged
MH  - Aged, 80 and over
MH  - Aggrecans/metabolism
MH  - Cartilage, Articular/*metabolism
MH  - Case-Control Studies
MH  - Chondrocytes/*metabolism
MH  - Collagen Type I/metabolism
MH  - Collagen Type II/metabolism
MH  - Comparative Genomic Hybridization
MH  - Female
MH  - Genes, Y-Linked
MH  - Humans
MH  - In Situ Hybridization, Fluorescence
MH  - Interleukin-1beta/metabolism
MH  - Male
MH  - Middle Aged
MH  - Osteoarthritis, Knee/*genetics/metabolism
MH  - RNA, Messenger/genetics
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - Sex Factors
MH  - Young Adult
EDAT- 2012/06/16 06:00
MHDA- 2012/12/28 06:00
CRDT- 2012/06/16 06:00
PHST- 2012/03/01 00:00 [received]
PHST- 2012/05/21 00:00 [revised]
PHST- 2012/05/30 00:00 [accepted]
PHST- 2012/06/16 06:00 [entrez]
PHST- 2012/06/16 06:00 [pubmed]
PHST- 2012/12/28 06:00 [medline]
AID - S1063-4584(12)00841-2 [pii]
AID - 10.1016/j.joca.2012.05.012 [doi]
PST - ppublish
SO  - Osteoarthritis Cartilage. 2012 Sep;20(9):1039-45. doi: 
      10.1016/j.joca.2012.05.012. Epub 2012 Jun 12.