Genomic chondrocyte culture profiling by array-CGH, interphase-FISH and RT-PCR

Stümm, Markus; Boger, Erich T.; Gaissmaier, Christoph; Oßwald, C.; Blankenburg, Markus; Wegner, Rolf-Dieter; Mollenhauer, J.

doi:10.1016/j.joca.2012.05.012

Cited by 9 publications

(18 citation statements)

References 21 publications

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“…In contrast, we observed a significantly lower IL-1b gene expression level after 3D cultivation as compared to monolayer cultivation (P < 0.05). Such increased IL-1b gene expression levels upon monolayer cultivation are uncommon and probably an effect of the diseased cartilage tissue harvested 27,28 . Eger et al reported higher IL-1b levels in osteoarthritic cartilage most likely being an expression of inflammatory activity.…”

Section: Discussionmentioning

confidence: 99%

Characterization of primary chondrocytes harvested from hips with femoroacetabular impingement

Bretschneider

Stiehler

Hartmann

et al. 2016

Osteoarthritis and Cartilage

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Section: Discussionmentioning

confidence: 99%

Characterization of primary chondrocytes harvested from hips with femoroacetabular impingement

Bretschneider

Stiehler

Hartmann

et al. 2016

Osteoarthritis and Cartilage

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“…Thus, it is vital that the risk of tumorigenic transformation due to growth factors’ use in vitro be examined. With increasing reports on the incidence of cytogenetic aberrations during cell ex vivo manipulation 28 , only a few attempts were made to investigate genetic alterations and tumorigenicity in chondrocytes induced by growth factors 29 . Therefore, in the current study, we investigated the biosafety of in vitro expanded human nasal septal chondrocytes (hNSCs) in growth factor cocktail supplemented culture medium.…”

Section: Introductionmentioning

confidence: 99%

Biosafety evaluation of culture-expanded human chondrocytes with growth factor cocktail: a preclinical study

Al-Masawa

Zaman

Chua

2020

Sci Rep

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The scarcity of chondrocytes is a major challenge for cartilage tissue engineering. Monolayer expansion is necessary to amplify the limited number of chondrocytes needed for clinical application. Growth factors are often added to improve monolayer culture conditions, promoting proliferation, and enhancing chondrogenesis. Limited knowledge on the biosafety of the cell products manipulated with growth factors in culture has driven this study to evaluate the impact of growth factor cocktail supplements in chondrocyte culture medium on chondrocyte genetic stability and tumorigenicity. The growth factors were basic fibroblast growth factor (b-FGF), transforming growth factor β2 (TGF β2), insulin-like growth factor 1 (IGF-1), insulin-transferrin-selenium (ITS), and platelet-derived growth factor (PD-GF). Nasal septal chondrocytes cultured in growth factor cocktail exhibited a significantly high proliferative capacity. Comet assay revealed no significant DNA damage. Flow cytometry showed chondrocytes were mostly at G0-G1 phase, exhibiting normal cell cycle profile with no aneuploidy. We observed a decreased tumour suppressor genes’ expression (p53, p21, pRB) and no TP53 mutations or tumour formation after 6 months of implantation in nude mice. Our data suggest growth factor cocktail has a low risk of inducing genotoxic and tumorigenic effects on chondrocytes up to passage 6 with 16.6 population doublings. This preclinical tumorigenicity and genetic instability evaluation is crucial for further clinical works.

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“…The lack of chromosomal aberrations in the healthy age-matched controls suggest that there is an association between chromosomal aberrations and osteoarthritis (Castellanos et al, 2004). According to Stumm et al (2012), the observed presence or absence of the Y chromosome does not appear to influence the transcriptional activity of chondrocytes.…”

Section: Gonosomal Chromosomesmentioning

confidence: 98%

“…PDLs. Finally, Stumm et al (2012), using human chondrocytes (articular cartilage tissue from patients with total knee replacement surgery and healthy cartilage from human organ donors) reveal autosomal stability, but losses of Y chromosomes.…”

Section: Autosomal Chromosomesmentioning

confidence: 99%

“…Single structural chromosomal aberrations in living donors scheduled for ACT include inversions, translocations, losses/gains of gonosomes, gains of chromosomes 5, 7 and 8 and marker chromosomes (Trimborn et al, 2012). Stumm et al (2012), in a study on genomic chondrocyte culture profiling by array-CGH and FISH, show autosomal stability but also variable loss of Y chromosome in all male samples in monolayer. Although losses of Y chromosomes may represent age-related processes, these data suggest some caution toward applying cultivation processes to chondrocytes from elderly patients for cell-based therapy.…”

Section: Introductionmentioning

confidence: 99%

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Comprehensive high-resolution genomic profiling and cytogenetics of human chondrocyte cultures by GTG-banding, locus-specific FISH, SKY and SNP array

Wallenborn¹,

Petters²,

Rudolf³

et al. 2018

eCM

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In the development of cell-based medicinal products, it is crucial to guarantee that the application of such an advanced therapy medicinal product (ATMP) is safe for the patients. The consensus of the European regulatory authorities is: "In conclusion, on the basis of the state of art, conventional karyotyping can be considered a valuable and useful technique to analyse chromosomal stability during preclinical studies". 408 chondrocyte samples (84 monolayers and 324 spheroids) from six patients were analysed using trypsin-Giemsa staining, spectral karyotyping and fluorescence in situ hybridisation, to evaluate the genetic stability of chondrocyte samples from non-clinical studies. Single nucleotide polymorphism (SNP) array analysis was performed on chondrocyte spheroids from five of the six donors. Applying this combination of techniques, the genetic analyses performed revealed no significant genetic instability until passage 3 in monolayer cells and interphase cells from spheroid cultures at different time points. Clonal occurrence of polyploid metaphases and endoreduplications were identified associated with prolonged cultivation time. Also, gonosomal losses were observed in chondrocyte spheroids, with increasing passage and duration of the differentiation phase. Interestingly, in one of the donors, chromosomal aberrations that are also described in extraskeletal myxoid chondrosarcoma were identified. The SNP array analysis exhibited chromosomal aberrations in two donors and copy neutral losses of heterozygosity regions in four donors. This study showed the necessity of combined genetic analyses at defined cultivation time points in quality studies within the field of cell therapy.

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Genomic chondrocyte culture profiling by array-CGH, interphase-FISH and RT-PCR

Cited by 9 publications

References 21 publications

Characterization of primary chondrocytes harvested from hips with femoroacetabular impingement

Characterization of primary chondrocytes harvested from hips with femoroacetabular impingement

Biosafety evaluation of culture-expanded human chondrocytes with growth factor cocktail: a preclinical study

Comprehensive high-resolution genomic profiling and cytogenetics of human chondrocyte cultures by GTG-banding, locus-specific FISH, SKY and SNP array

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