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Genomic chondrocyte culture profiling by array-CGH, interphase-FISH and RT-PCR.
M Stumm1, E Boger, C G Gaissmaier
1BG Berlin-Genetics GmbH, MDC-Buch, Berlin, Germany. stumm@kudamm-199.de
Osteoarthritis and Cartilage
|June 16, 2012
Summary
Cultured chondrocytes from male donors may lose their Y-chromosome with age, but this does not affect cell function. Caution is advised when using chondrocytes from older male patients for tissue engineering.
Area of Science:
- Cell Biology
- Genomics
- Tissue Engineering
Background:
- In vitro expansion of chondrocytes is crucial for cartilage tissue engineering.
- Prolonged cell culture may induce genomic alterations.
- Understanding these changes is vital for clinical applications.
Purpose of the Study:
- To investigate in vitro induced genomic changes in human chondrocytes.
- To assess the impact of cell expansion on chondrocyte genomes.
- To evaluate potential functional consequences of genomic alterations.
Main Methods:
- Array comparative genomic hybridization (aCGH) and fluorescence in situ hybridization (FISH) were used for molecular karyotyping.
- Analysis of DNA from 25 chondrocyte samples (9 donors) and 28 samples (16 individuals).
- Reverse-transciptase polymerase chain reaction (RT-PCR) assessed chondrocytic mRNA phenotype.
Main Results:
- Autosomal stability was observed in all samples.
- Male samples showed variable loss of the Y-chromosome, potentially age-dependent.
- No correlation between Y-chromosome loss and mRNA expression of collagen types I, II, aggrecan, or IL-1ß was found.
Conclusions:
- Cultured chondrocytes from male donors may experience age-related Y-chromosome loss.
- This loss did not appear to cause functional impairment in the studied chondrocytes.
- Caution is recommended when using chondrocytes from elderly male patients in tissue engineering.
