Establishment of Cerebral Organoid‐like Structures from Dental Pulp Stem Cells (DPSCs) to Model Neurodegenerative Diseases.

Background 3D-cell cultures are closer to in vivo conditions than the monolayers. There different culture models, most relevant is Cerebral Organoids (CO) model (Lancaster et al. 2014), it can show complex processes and proved molecular structural changes when modeling Alzheimer´s disease (AD), other diseases. One of main limitations using this requirement for Induced Pluripotent Stem Cells (iPSC) or Human Embryonic (hESC) CO generation; because iPSC could undergo epigenetic during process reprogramming; moreover, hESC controversial their ethical implications. We proposed use DPSCs instead generate organoid-like structures due capability differentiate into neuronal lineage, lack pluripotency, its easy access. Method The were obtained by dissociating dental pulp a healthy 46 years old female with colagenase-dispase mix at 37°C, isolated cultured. cells phenotype was characterized Western-Blot, RT-PCR immunofluorescence. cultured DPSC low-adherence wells 6 days DMEM/F12 medium + thiazovivin 50 mM, bFGF 4ng/mL, BFS 20%, Glutamax® 200x, MEM-NEAA 1:100, 2-mercaptoethanol 100 µm murine LIF 10 µg/mL until cell aggregation embryoid-like structures. After that, neural induction carried out. Result successfully detecting pluripotential markers OCT4, Actin, NanoG, mesenchymal CD105, CD73, marker β-III Tubulin which always present as well hematopoietic CD14, CD34, CD45. Organoids-like from de after 35 differentiation protocol detected NeuN, MAP2, Tubulin, tau, all that claim phenotype. Conclusion Our results suggest potential approach generation benefits shorter time less expensive likewise, resemble aging changes. Further studies needed validate neurodegenerative diseases such AD.