TITRE: Trebuchet MS, 40pt, white Authors: Arial, 20pt 1. Affiliation 1 CNRS, Univ Paris Diderot, Sorbonne Paris Cité, École nationale vétérinaire d'Alfort,

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1 TITRE: Trebuchet MS, 40pt, white Authors: Arial, 20pt 1. Affiliation 1 CNRS, Univ Paris Diderot, Sorbonne Paris Cité, École nationale vétérinaire d'Alfort, F- 75010, Paris, France 2. Affiliation 2 Descriptif Logo collabor ators

2 Titre: Trebuchet MS, 28pt, gras, bleu foncé IntroductionMethodsResultsDiscussion  Arial, 20pt, bleu foncé (R21/G68/B101) ou noir Liste/Conclusion  Puces carrées, Rouge (R210/G2/B60)  1.Author et al. Year. J. Acronym 2.Petite et al. 2012. Nat Biotech

3 Titre: Trebuchet MS, 28pt, gras, bleu foncé IntroductionMethodsResultsDiscussion  Arial, 20pt, bleu foncé (R21/G68/B101) ou noir Liste/Conclusion  Puces carrées, Rouge (R210/G2/B60)  1.Author et al. Year. J. Acronym 2.Petite et al. 2012. Nat Biotech

4 Titre: Trebuchet MS, 28pt, gras, bleu foncé IntroductionMethodsResultsDiscussion  Arial, 20pt, bleu foncé (R21/G68/B101) ou noir Liste/Conclusion  Puces carrées, Rouge (R210/G2/B60)  1.Author et al. Year. J. Acronym 2.Petite et al. 2012. Nat Biotech

5 Titre: Trebuchet MS, 28pt, gras, bleu foncé IntroductionMethodsResultsDiscussion  Arial, 20pt, bleu foncé (R21/G68/B101) ou noir Liste/Conclusion  Puces carrées, Rouge (R210/G2/B60)  1.Author et al. Year. J. Acronym 2.Petite et al. 2012. Nat Biotech This work was supported by X institution (grant reference) Logo founding institutes

6 Mimicking developmental cell biology to regenerate the intervertebral disc Esther POTIER 1, K. ITO 2, S. de VRIES 2, I. ARKESTEIJN 2 1. UMR 7052 Osteo-Articular Bioengineering and Bioimaging CNRS, Univ Paris Diderot, Sorbonne Paris Cité, École nationale vétérinaire d'Alfort, F- 75010, Paris, France 2. Eindhoven University of Technology Department of Biomedical Engineering, Eindhoven, The Netherlands

7 Low back pain IntroductionMethodsResultsDiscussion  Cost: 100-200 billions/year in USA only 1  In 40% of cases: intervertebral disc degeneration involved 2 1. Katz. 2006. JBJS Am 2. Freemont. 2009. Rheum

8 Direct long-term co-culture IntroductionMethodsResultsDiscussion BMSC+NPC 100 NC 0 BMSC+NPC 100 NC 25 BMSC+NPC 100 NC 50 4 weeks hgDMEM + 10%FBS + 1%P/S at 2% O 2  Groups  Culture  Analysis Cell morphology (confocal) and group composition (+FACS) Matrix deposition (GAG, HYP, Alcian Blue) Cell phenotype (RT-qPCR for col2, agg, T, CK8/18/19)

9 Direct long-term co-culture IntroductionMethodsResultsDiscussion  No stimulation of matrix production or discogenic phenotype Matrix production at day 28. Means ± standard deviation; N = 4; *p < 0.05 vs. BMSC-Ch; § p < 0.05 vs. 1.5SF or 1.5NC  Lose of NC phenotype during culture  FBS in medium? NC phenotype at day 28. Means ± standard deviation; N = 4; *p < 0.05 vs. fresh native NCs

10 NC conditioned medium IntroductionMethodsResultsDiscussion  NPCs can be stimulated by: BMSCs NCCM (-) High number of cells needed (1:1) (-) Cell harvesting and expansion (+) Can use higher concentration (+) Can identify molecular factor(s) involved (-) Sustain release