REGENERATION OF THE INTERVERTEBRAL DISC (IVD) USING IN VITRO DIFFERENTIATED STEM CELLS

• Published in: International journal of artificial organs 2011-08, Vol.34 (8), p.690-691
• Main Authors: Ehlicke, F, Freimark, D, Pudlas, M, Dorresteijn, A, Walles, H, Czermak, P
• Format: Article
• Language: English
• Subjects: Chondrocytes
• ISSN: 0391-3988

Objectives: The degeneration of the nucleus pulposus (NP) is one major reason for low back pain. One possible method of treatment is cell-based therapy with differentiated human mesenchymal stem cells (hMSC). For a differentiation into NP cells, the hMSCs require a 3D environment and various stimuli such as growth factors. In our work, we want to identify the optimal stimulation for the differentiation of hMSCs into NP cells. Regarding the therapeutical use it is indispensable to verify the differentiation success of hMSCs into NP cells and to delimit the differentiated cells to chondrocytes. As Raman spectroscopy has a high potential for non-invasive characterization of suspension cells and distinction between different cell lines, our research group together with the Fraunhofer Institute of Interfacial Engineering and Biotechnology (IGB) tested the applicability of this method for cells embedded in hydrogels. Methods: hMSCs were cultivated three-dimensionally to form NP cells. To investigate the putative differentiation stimulating ability of several growth factors and components of the extracellular matrix (ECM), RT-PCR as well as fluorescence immunostaining of NP-speciffc marker proteins were done. Results: In all differentiation experiments with growth factors, NP-specific marker proteins were expressed. Data concerning the differentiation of hMSC under the influence of ECM components will be presented. Using Raman Spectroscopy as well as common methods of molecular biology, we were able to distinguish NP cells and differentiated hMSC from undifferentiated stem cells and chondrocytes. Conclusions: The expression of NP-specific marker proteins indicates the ability of three growth factors to differentiate hMSCs into NP-like cells. Using Raman spectroscopy and RT-PCR we could clearly display that NP cells differ from chondrocytes. As only pure NP cells (e.g. into NP cells differentiated hMSCs) could be used for subsequent therapeutical use, this finding is of great benefit for NP regeneration approaches.