eCM (Eur Cell Mater / e Cells & Materials) Not-for-profit Open Access
Created by Scientists, for Scientists
 ISSN:1473-2262         NLM:100973416 (link)         DOI:10.22203/eCM

2018   Volume No 35 – pages 225-241

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

Authors: M Wallenborn, O Petters, D Rudolf, H Hantmann, M Richter, P Ahnert, L Rohani, JJ Smink, GC Bulwin, W Krupp, RM Schulz, H Holland

Address: Saxonian Incubator for Clinical Translation (SIKT) and Faculty of Medicine, University of Leipzig, D-04103 Leipzig, Germany

E-mail: Heidrun.Holland at medizin.uni-leipzig.de


Abstract: 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.

Key Words: ATMP, CBMP, cell therapy, chondrocyte, GTG-banding, spectral karyotyping, SNP array.

Publication date: April 23rd 2018

Article download: Pages 225-241 (PDF file)
DOI:
10.22203/eCM.v035a16

This Article contains the following errors:

1. “To obtain an EU-wide marketing authorisation, in 2013, Barkholt et al. performed further investigations — particularly regarding safety, including cellular stability — on chondrocyte samples, to rule out any side effect and risk of the treatment method, as requested by the EMA.” should read “To obtain an EU-wide marketing authorisation, in 2015, Zscharnack et al. performed further investigations — particularly regarding safety, including cellular stability — on chondrocyte samples, to rule out any side effect and risk of the treatment method, as requested by the EMA.”

2. Spherox®" should read "chondrocyte samples from non-clinical studies".


The study was performed as part of the non-clinical development, from 2013 until 2015, in the context of the approval process for the Advanced Therapy Medicinal Product (ATMP) Spherox of the CO.DON AG, which was approved by the EC/EMA in July 2017. The product Spherox itself has not been subjected to any of the described analyses. The objects of our investigation were chondrocyte samples from non-clinical studies carried out as part of the approval process for Spherox.

 

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