Abstract

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.

Links

Publisher Full Text

Authors

Stumm M, Boger E, Gaissmaier CG, Oßwald C, Blankenburg M, Wegner RD, Mollenhauer JA

Institution

BG Berlin-Genetics GmbH, MDC-Buch, Berlin, Germany. stumm@kudamm-199.de

Source

Osteoarthritis and cartilage / OARS, Osteoarthritis Research Society 20:9 2012 Sep pg 1039-45

MeSH

Adult
Aged
Aged, 80 and over
Aggrecans
Cartilage, Articular
Case-Control Studies
Chondrocytes
Collagen Type I
Collagen Type II
Comparative Genomic Hybridization
Female
Genes, Y-Linked
Humans
In Situ Hybridization, Fluorescence
Interleukin-1beta
Male
Middle Aged
Osteoarthritis, Knee
RNA, Messenger
Reverse Transcriptase Polymerase Chain Reaction
Sex Factors
Young Adult

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

22698443