Enriching Stem/Progenitor Cells from Dental Pulp Cells by Low-density Culturing

TY - JOUR

T1 - Enriching Stem/Progenitor Cells from Dental Pulp Cells by Low-density Culturing

AU - Zhao, Yao

AU - Zheng, Yajie

AU - Eichhorn, Wolfgang

AU - Klatt, Jan

AU - Henningsen, Anders

AU - Kluwe, Lan

AU - Friedrich, Reinhard E

AU - Gosau, Martin

AU - Smeets, Ralf

N1 - Copyright© 2019, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.

PY - 2018/12/28

Y1 - 2018/12/28

N2 - BACKGROUND/AIM: Clonogenicity is a key feature of stem/progenitor cells. The present study aimed to enrich stem/progenitor cells from dental pulp cells by means of culturing the cells at a low clonal density with spatial separation and the evaluate differentiation potential of the surviving cells.MATERIALS AND METHODS: Pulp cells derived from wisdom teeth were seeded into wells of a 96-plate at a mean density of 1 cell/well and cultured for 2 weeks. Surviving cells were harvested, pooled together and subjected to differentiation into adipocytes, osteoblasts and neurons using respective inducing conditions for 3 weeks. The former two types of cells were examined by staining with Oil Red O and Alizarin Red, respectively. Neuron-like cells were inspected for their morphology and immunostained for microtubule-associated protein 2 and β-tubulin III.RESULTS: Vital cells were obtained in eight wells of a 96-well plate, corresponding to a survival rate of 8%. Since fewer than two wells would be expected to contain more than four cells at seeding, the majority of surviving cells likely grew from 1-3 cells, which is a very low density. These cells differentiated into functional adipocytes and osteoblasts, and morphologically neuron-like cells.CONCLUSION: Low-density seeding with spatial separation enables statistical estimation of cell number in wells and provides an effective strategy for enriching stem/progenitor cells and for isolating clonal dental pulp cells.

AB - BACKGROUND/AIM: Clonogenicity is a key feature of stem/progenitor cells. The present study aimed to enrich stem/progenitor cells from dental pulp cells by means of culturing the cells at a low clonal density with spatial separation and the evaluate differentiation potential of the surviving cells.MATERIALS AND METHODS: Pulp cells derived from wisdom teeth were seeded into wells of a 96-plate at a mean density of 1 cell/well and cultured for 2 weeks. Surviving cells were harvested, pooled together and subjected to differentiation into adipocytes, osteoblasts and neurons using respective inducing conditions for 3 weeks. The former two types of cells were examined by staining with Oil Red O and Alizarin Red, respectively. Neuron-like cells were inspected for their morphology and immunostained for microtubule-associated protein 2 and β-tubulin III.RESULTS: Vital cells were obtained in eight wells of a 96-well plate, corresponding to a survival rate of 8%. Since fewer than two wells would be expected to contain more than four cells at seeding, the majority of surviving cells likely grew from 1-3 cells, which is a very low density. These cells differentiated into functional adipocytes and osteoblasts, and morphologically neuron-like cells.CONCLUSION: Low-density seeding with spatial separation enables statistical estimation of cell number in wells and provides an effective strategy for enriching stem/progenitor cells and for isolating clonal dental pulp cells.

KW - Adipocytes/cytology

KW - Cell Culture Techniques

KW - Cell Differentiation/genetics

KW - Cell Separation

KW - Dental Pulp/cytology

KW - Humans

KW - Mesenchymal Stem Cells/cytology

KW - Osteoblasts/cytology

KW - Stem Cells/cytology

U2 - 10.21873/invivo.11434

DO - 10.21873/invivo.11434

M3 - SCORING: Journal article

C2 - 30587598

VL - 33

SP - 23

EP - 29

JO - IN VIVO

JF - IN VIVO

SN - 0258-851X

IS - 1

ER -