(149) PD.SPHERE, a Novel Three-dimension Cellular Model for Peyronie’s Disease via ‘Microgravity’ Condition: The New Era of in vitro Cell Culture-based Model for PD
Notice bibliographique
Résumé
Abstract Introduction Peyronie's disease (PD) is a common problem that affects 4-13% of the male population and results in a significant decline in patient and partner quality of life. Currently, in vitro monolayer fibroblast culture and animal models are actively used to study pathogenesis and treatments. However, both models are not a great pathophysiological representative for human PD due to the lack of cellular architecture and limited insight into the PD mechanism. Objective We aimed to develop a three-dimensional (3D) cellular model to reveal the PD pathogenesis mechanism and perhaps use this model as a treatment screening tool. Methods Human PD fibroblasts isolated from PD patients during surgery were cultured in various non-adhesive culture plates under the 'microgravity' condition. Complete media (DMEM+10% FBS+2mM L-glutamine) with and without TGF-β supplement was used to optimize the conditions for the PD.SPHERE, a 3D cell culture model of PD fibroblasts. Molecular probes against nucleus, actin, vimentin (VIM) and smooth muscle actin (SMA) were used to study the morphology and expression of molecular markers. Confocal-microscopic analyses were performed to obtain the morphological snapshots of and molecular data of the PD.SPHERE. Results We successfully developed a 3D cell culture (spheroid) model from human PD fibroblasts with our defined conditions. The PD spheroids were formed individually in each vessel of a 96-well plate coated with a non-adhesive substance. A PD.SPHERE was made in each well in a 96-well plate from three different individuals. Moreover, the size and circularity of spheroids were consistent across the plate. Although TGF-β supplemented condition significantly increased the cell division capability of the PD fibroblasts in the monolayer model, this supplementation was not necessary for our 3D model since TGF-β did not significantly increase the size and circularity of the 3D model. Despite the spheroids' increases in cell density over the culturing period, TGF-β did not affect the cell density in the 3D-cellular structure. Throughout the PD.SPHERE architecture, it also expressed SMA and VIM, the intracellular skeletal markers. Conclusions In conclusion, our model is consistent, reproducible and morphologically relevant to a PD plaque. Our model also demonstrates that we have the potential to produce high throughput treatment screening platforms for PD. Additionally, the PD.SPHERE platform is personalized and customizable to the specific individual; thus, our model indicated the potential to develop this platform for personalized treatment planning. However, signature gene expression and physiological and immunological information (i.e., collagen production, extracellular matrix organization, cellular expression of tight junctions, chemokines, and cytokines) should be further collected to verify the pathophysiological, immunological and treatment response for this model. Disclosure No
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Comment cette classification a été obtenuedéplier
Prédiction distillée sur la base complète
Imitation des enseignantsNi prévalence calibrée, ni vérité terrain. Validation humaine à venir. Apprise à partir de 10 348 étiquettes directes de Codex et de 10 348 étiquettes directes de Gemma. Le mode candidate est l'union des têtes enseignantes seuillées; le consensus est leur intersection. Ces sorties portent le statut machine_predicted_unvalidated et ne sont ni des étiquettes humaines ni des étiquettes directes de modèles de pointe.
Scores Codex et Gemma par catégorie
| Catégorie | Codex | Gemma |
|---|---|---|
| Métarecherche | 0,001 | 0,000 |
| Méta-épidémiologie (sens strict) | 0,000 | 0,000 |
| Méta-épidémiologie (sens large) | 0,001 | 0,000 |
| Bibliométrie | 0,000 | 0,000 |
| Études des sciences et des technologies | 0,000 | 0,000 |
| Communication savante | 0,000 | 0,000 |
| Science ouverte | 0,000 | 0,000 |
| Intégrité de la recherche | 0,000 | 0,000 |
| Charge utile insuffisante (le modèle a refusé de juger) | 0,000 | 0,000 |
Scores machine (provisoires)
Les deux têtes enseignantes du modèle étudiant, lues sur ce travail. Un score ordonne la base pour la relecture; il n'affirme jamais une catégorie, et le statut de validation accompagne chaque rangée tel quel.
Scores de référence d'un modèle non mature (critères de maturité non atteints, 7 itérations). Un score ordonne; il n'affirme jamais une catégorie.
score_only:v0-immature-baseline · tel quel depuis la passe de notation : score_only signifie que le nombre peut ordonner les travaux, et qu'aucune étiquette de catégorie n'en découleClassification
machine, non validéePrédiction automatique; un appel candidat d’une seule tête enseignante, pas un consensus.
Le détail, modèle par modèle et score par score, se trouve en fin de page sous « Comment cette classification a été obtenue ».