gms | German Medical Science

Deutscher Kongress für Orthopädie und Unfallchirurgie (DKOU 2016)

25.10. - 28.10.2016, Berlin

GMP-compliant production of a mesenchymal stem cell based cartilage therapy

Meeting Abstract

  • presenting/speaker Christian Schmidt - Biotechnologisch-Biomedizinisches Zentrum , Leipzig, Germany
  • Oliver Petters - Biotechnologisch-Biomedizinisches Zentrum, Leipzig, Germany
  • Gero Hütter - Cellex, Dresden, Germany
  • Bastian Marquaß - Universität Leipzig, Klinik für Orthopädie, Unfallchirurgie und Plast. Chirurgie, Leipzig, Germany
  • Ralf Henkelmann - Universität Leipzig, Klinik für Orthopädie, Unfallchirurgie und Plast. Chirurgie, Leipzig, Germany
  • Ronny Schulz - Biotechnologisch-Biomedizinisches Zentrum, Leipzig, Germany

Deutscher Kongress für Orthopädie und Unfallchirurgie (DKOU 2016). Berlin, 25.-28.10.2016. Düsseldorf: German Medical Science GMS Publishing House; 2016. DocPO29-1179

doi: 10.3205/16dkou779, urn:nbn:de:0183-16dkou7794

Veröffentlicht: 10. Oktober 2016

© 2016 Schmidt et al.
Dieser Artikel ist ein Open-Access-Artikel und steht unter den Lizenzbedingungen der Creative Commons Attribution 4.0 License (Namensnennung). Lizenz-Angaben siehe



Objectives: Due to the limited regeneration potential of articular cartilage, it is still an ambitious clinical challenge to treat cartilage defects with long-term success. Current treatment modalities like microfracture and (matrix-associated) autologous chondrocyte transplantation (M/ACT) have several disadvantages including formation of inferior fibrocartilage tissue and chondrocyte de-differentiation.

Alternatively, application of mesenchymal stromal cells (MSCs) instead of chondrocytes in MACT-like grafts has shown a beneficial outcome compared to chondrocyte-based MACT in large animal models. However, one of the main challenges is the translation from the basic research approach to a clinical applicable MSC-based MACT graft under consideration of the regulatory framework.

Here, we evaluated the prospective GMP production process of a MSC-MACT implant in terms of the logistics of human bone marrow (hBM), as well as cell isolation, expansion and differentiation of the MSCs.

Methods: HBM samples of the iliac crest and GMP-compliant raw materials were used to evaluate various process variables. This includes the investigation of different storage and transport conditions, various density gradients for mononuclear cells (MNC) isolation and different expansion media as well as differentiation protocols within a 3D collagen matrix. The process parameters were examined with respect to the following quality control aspects: cell viability, apoptosis, necrosis, frequency, senescence, proliferation, number of MNC, CFU-F and MSC, cell identity, purity and genetic stability.

Results and Conclusion: The immediate processing of hBM was beneficial concerning the quality of MSC compared to longer transport conditions. The logistics at room temperature instead at 4°C showed advantageous properties regarding the yield and quality of MSC.

Although, the isolation of MNC is slightly increased using a higher density gradient of 1.084 mg/ml compared to 1.073 and 1.077 mg/ml, there was no significant differences regarding the yield of MNCs, number of CFUs, immunophenotype, proliferation and differentiation behavior.

While the yield of MSC and their identity differs remarkably, no significant changes in CFU-F assay were observed between the 9 different GMP expansion media variations. Depending on the media, the chondrogenic differentiation is altered.

Both the sequence and the administered dose/s of TGF-β3 affect the chondrogenesis of the expanded MSCs in 3D matrix massively.

With this translational in vitro study it becomes evident that every single process variable, especially the logistic conditions of the starting material and the applied expansion media, substantially influences the product quality. The optimized production process of such an MACT-like MSC implant fulfills the ISCT recommendations regarding the quality, cell yield, identity and purity. In summary the GMP-compliant production of a MSC-MACT is feasible.