GP16: The effects of 3D culture on the expansion and maintenance of nucleus pulposus progenitor cell multipotency

Guerrero, Julien; Häckel, Sonja; Croft, Andreas S.; Albers, Christoph E.; Gantenbein, B. (2021). GP16: The effects of 3D culture on the expansion and maintenance of nucleus pulposus progenitor cell multipotency (Unpublished). In: ISSLS Virtual Meeting. Milano. 2-4 June 2021.

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Low back pain (LBP) is a global health concern that affects as many as 75–80% of people during their lifetime (1). The cause of LBP is multifactorial; however, intervertebral disc (IVD) degeneration is a major factor (2). Therefore, stem cell therapy to activate self-repair of the IVD appears to be an exciting treatment strategy (3). In this respect, tissue-specific progenitors may play a crucial role for IVD regeneration, as these cells are perfectly adapted to the IVD niche. Such a rare progenitor cell population residing in the nucleus pulposus (NP) was found to be positive for the angiopoietin-1 receptor (Tie2+) and was demonstrated to possess self-renewal capacity and in vitro multipotency (4). However, extremely low numbers of Tie2+ cells in NP limits the feasibility of potential cell therapy strategies. In this study, the first aim was to optimize culture conditions to mimic NP microenvironment for Tie2+ cells expansion. The second aim was to study the influence of cells expansion in various conditions on the chondrogenic potential of those cells.
Here, cells were obtained from NP tissue of IVD in vertebrae of donors undergoing spinal surgery with written consent. Briefly, after mild overnight digestion of the NP tissue cells were cultivated within different conditions (in 2D or 3D) and culture media (fibronectin coating, TGFβ-1 and/or FGF-2) to mimic the microenvironment and the niche of the native NP of IVD. Subsequently, to study their regenerative potential cells were seeded into a 3D hydrogel scaffold collagen based and induced into the chondrogenic lineage for 2-3 weeks and analyzed.
The present study proposes to exploit physiological niches in order to better maintain the functionality of cells during their in vitro expansion. This project not only has a scientific impact by evaluating the role of native physiological niches on the functionality of NP progenitors but could also lead to an innovative clinical approach with cell therapy for IVD regeneration and repair in a context of LBP.
Acknowledgements: Financial support was received from iPSpine H2020 project #825925.
References: (1) – Hartvigsen et al., 2018, Lancet; (2) – Martirosyan et al., 2016, Frontiers in Surgery; (3) – Sakai et al., 2015, Nature Reviews, Rheumathology; (4) – Frauchiger et al., 2019, Tissue Engineering. Part C, Methods.

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