Poster De Conférence Année : 2024

First co-culture of human dural fibroblasts and adipose stem cells for dura mater tissue-engineering

Résumé

Introduction The dura mater, the meninges’ outermost layer, acts as a semi-rigid barrier between the central nervous system and bone tissues [1]. Its limited healing ability and tendency to form scar tissue can lead to complications after neurosurgery and dural resection. Advancements in dural tissue engineering could enhance our understanding of this interface and help create in vitro models and implantable scaffolds for dura mater repair. One challenge is to develop a co-culture model using human dural fibroblasts (HDufs) and adipose-derived stem cells (ASC52telo) differentiated into osteoblasts, aiming to mimic natural tissue environments and study cell interactions. Here, we present the phenotype analysis results for HDufs and ASC52telo after up to one month of co-culture. Methods Two co-culture methods, indirect (separate compartments) and direct (mixed), were conducted using HDufs and ASC52telo. Initially, cells were cultured in an optimized proliferation medium for one week. Then, a three-week period followed with or without osteogenic factors (dexamethasone, ascorbic acid, and beta-glycerophosphate). To assess the effects of co-culture methods and osteogenic stimulation on the behavior of HDufs and ASC52telo, qPCR, immuno-fluorescence, ALP staining, and alizarin red staining were performed at 14 and 28 days. Monoculture controls were conducted to validate the study. Results and discussion The results revealed increased expression of osteogenic genes and bone biomarkers during co-culture, indicating stem cell maturation into osteoblasts even without differentiation factors, suggesting a beneficial impact of HDufs interaction on this process. Regarding HDufs' functionality in long-term co-culture, they exhibited a notable phenotype change: increased expression of dura mater periosteal layer-related markers and Crapb2 decrease, specific of the dural border cell layer [2] (native dura deepest layer). The dural fibroblast marker FxyD5 was expressed all along till the co-culture end. Conclusion Long-term co-culture of HDufs and ASC52telo proved stable, ensuring survival and functionality of both cell types. The synergy between them facilitated stem cell differentiation into mature osteoblasts while inducing phenotypic changes in HDufs resembling native tissue. These findings support the potential of this co-culture model for in vitro dura mater studies and as a substitute testing model for in vivo purposes. Next, we plan to validate these results with 3D scaffold co-culture. References 1. Vandenabeele et al, J Anat, 189:417–30, 1996. 2. Farmer and al.,Nat Commun, 12:4797, 2021

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hal-05039368 , version 1 (18-04-2025)

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  • HAL Id : hal-05039368 , version 1

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Jean-Philippe Tosiani, Nathalia Oderich Muniz, Timothée Baudequin. First co-culture of human dural fibroblasts and adipose stem cells for dura mater tissue-engineering. 50th congress of the European Society for Artificial Organs, Sep 2024, Aachen (Aix la Chapelle), Germany. The International Journal of Artificial Organs. ⟨hal-05039368⟩
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