Optimizing Conditions for Spheroid Formation of Dental Pulp Cells in Cell Culture
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Optimizing Conditions for Spheroid Formation of Dental Pulp Cells in Cell Culture. / Zheng, Yajie; Jiang, L I; Yan, Ming; Gosau, Martin; Smeets, Ralf; Kluwe, Lan; Friedrich, Reinhard E.
in: IN VIVO, Jahrgang 35, Nr. 4, 29.06.2021, S. 1965-1972.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
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TY - JOUR
T1 - Optimizing Conditions for Spheroid Formation of Dental Pulp Cells in Cell Culture
AU - Zheng, Yajie
AU - Jiang, L I
AU - Yan, Ming
AU - Gosau, Martin
AU - Smeets, Ralf
AU - Kluwe, Lan
AU - Friedrich, Reinhard E
N1 - Copyright © 2021 International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.
PY - 2021/6/29
Y1 - 2021/6/29
N2 - BACKGROUND/AIM: Spheroid formation is a well-known feature of stem/progenitor cells. Dental pulp cells (DPCs) cultured in serum-free medium can also form spheroids. However, the success rate varies largely depending on various factors. This study aimed to explore these factors and optimize the conditions.MATERIALS AND METHODS: Primary DPCs were obtained from 6 wisdom teeth. Possible influencing factors including donor teeth, concentrations of the KnockOut Serum Replacement (KSR), seeding density (regarding surface and volume), passage and freezing were tested.RESULTS: DPCs from all 6 donor teeth formed spheroids in serum-free medium. Number, size, and total area of spheroids varied among different donor teeth. Optimal concentration of the KSR and seeding densities also varied from tooth to tooth. Generally, high KSR and high cell density lead to better spheroid formation. However, very high KSR and cell density can also lead to cell death and the fusion of spheroids to irregular aggregates.CONCLUSION: An initial setting can be recommended as: Serum-free MEM plus 10-15% KSR and seeding densities of 1-2×105 cells/ml and 2×105 cells/cm2 These parameters provide a direction for optimizing the condition for obtaining spheroids from human DPCs.
AB - BACKGROUND/AIM: Spheroid formation is a well-known feature of stem/progenitor cells. Dental pulp cells (DPCs) cultured in serum-free medium can also form spheroids. However, the success rate varies largely depending on various factors. This study aimed to explore these factors and optimize the conditions.MATERIALS AND METHODS: Primary DPCs were obtained from 6 wisdom teeth. Possible influencing factors including donor teeth, concentrations of the KnockOut Serum Replacement (KSR), seeding density (regarding surface and volume), passage and freezing were tested.RESULTS: DPCs from all 6 donor teeth formed spheroids in serum-free medium. Number, size, and total area of spheroids varied among different donor teeth. Optimal concentration of the KSR and seeding densities also varied from tooth to tooth. Generally, high KSR and high cell density lead to better spheroid formation. However, very high KSR and cell density can also lead to cell death and the fusion of spheroids to irregular aggregates.CONCLUSION: An initial setting can be recommended as: Serum-free MEM plus 10-15% KSR and seeding densities of 1-2×105 cells/ml and 2×105 cells/cm2 These parameters provide a direction for optimizing the condition for obtaining spheroids from human DPCs.
KW - Cell Culture Techniques
KW - Cell Differentiation
KW - Cells, Cultured
KW - Culture Media
KW - Dental Pulp
KW - Humans
KW - Stem Cells
U2 - 10.21873/invivo.12464
DO - 10.21873/invivo.12464
M3 - SCORING: Journal article
C2 - 34182470
VL - 35
SP - 1965
EP - 1972
JO - IN VIVO
JF - IN VIVO
SN - 0258-851X
IS - 4
ER -
