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Optimizing Conditions for Spheroid Formation of Dental Pulp Cells in Cell Culture
Yajie Zheng1, L I Jiang1, Ming Yan1
1Department of Oral and Craniomaxillofacial Surgery, University Hospital Hamburg-Eppendorf, Hamburg, Germany.
In Vivo (Athens, Greece)
|June 28, 2021
Summary
Dental pulp cells (DPCs) can form spheroids in serum-free conditions, but success varies. This study identified optimal KnockOut Serum Replacement (KSR) concentrations and seeding densities for consistent spheroid formation from human DPCs.
Area of Science:
- Stem cell biology
- Tissue engineering
- Dental research
Background:
- Spheroid formation is characteristic of stem/progenitor cells.
- Dental pulp cells (DPCs) can form spheroids in serum-free medium.
- Spheroid formation success in DPCs is variable and influenced by multiple factors.
Purpose of the Study:
- To investigate factors influencing dental pulp cell (DPC) spheroid formation.
- To optimize culture conditions for consistent DPC spheroid generation.
- To establish reproducible protocols for DPC spheroid culture.
Main Methods:
- Primary DPCs were isolated from 6 human wisdom teeth.
- Cultured DPCs in serum-free medium with varying KnockOut Serum Replacement (KSR) concentrations.
- Tested different seeding densities (cell/volume and cell/surface area), passage numbers, and freezing conditions.
Main Results:
- DPCs from all donors formed spheroids, but characteristics varied by donor.
- Optimal KSR concentrations and seeding densities differed between donor teeth.
- Higher KSR and cell densities generally improved spheroid formation, but excessive levels caused cell death and aggregation.
Conclusions:
- Recommended initial settings: serum-free MEM with 10-15% KSR.
- Suggested seeding densities: 1-2x10^5 cells/ml and 2x10^5 cells/cm^2.
- These parameters provide a guideline for optimizing human DPC spheroid formation.
