Three dimensional culture of dental pulp cells and signal network analysis for clinical applications

2015 Fiscal Year Final Research Report

• Project/Area Number: 26893074
• Research Category: Grant-in-Aid for Research Activity Start-up
• Allocation Type: Single-year Grants
• Research Field: Dental engineering/Regenerative dentistry
• Research Institution: Tokyo Medical and Dental University
• Principal Investigator: Yamamoto Mioko 東京医科歯科大学, 歯学部附属病院, 医員 (50732749)
• Project Period (FY): 2014-08-29 – 2016-03-31
• Keywords: 三次元スフェロイド培養

Outline of Final Research Achievements

For 3D spheroid culture, mouse dental papilla cells (MDPs) were cultured for 24 hours in 96-well low-attachment culture plates. Flat-bottomed plates were used for two-dimensional (2D) monolayer culture as a control. Following RNA extraction and complementary DNA synthesis from samples, real-time PCR was performed to evaluate the expression of alkaline phosphatase (Alp) and bone morphogenetic protein2 (BMP2) using specific primers. Up-regulation of Alp and Bmp2 expression was observed in the 3D spheroid-cultured MDPs, however, the expression of these genes was low in the 2D monolayer-cultured MDPs. In order to study the effects of integrin signaling, Focal adhesion kinase and Extracellular Signal-regulated Kinase 1/2 inhibitors were added to the 3D-spheroid cultured MDPs. Then, down-regulation of Alp and Bmp2 was observed in the presence of these inhibitors. In conclusion, 3D-spheroid culture induced the odonto-/osteoblastic differentiation of MDPs probably through integrin signaling.

Free Research Field

歯内療法