Characterization of mesenchymal progenitor cells in crown and root pulp from human mesiodentes

Sato, Momoko; Toriumi, Taku; Watanabe, Nobuyuki; Watanabe, Eri; Akita, Daisuke; Mashimo, Takayuki; Akiyama, Yoshitsugu; Isokawa, Keitaro; Shirakawa, Tetsuo; Honda, MJ

doi:10.1111/odi.12234

Cited by 11 publications

(7 citation statements)

References 38 publications

(56 reference statements)

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“…Migrated cells were plated at 1 × 10 5 cells per well in 6-well plates and cultured for 1, 3, 5, or 7 days in neural induction medium or mesenchymal stem cell (MSC) growth medium consisting of the alpha modification of Eagle's medium (α-MEM; Wako) supplemented with 10% fetal bovine serum, 50 U/mL penicillin, and 50 μg/mL streptomycin (Wako). Cell proliferation was determined by water-soluble tetrazolium salt (WST)-8 assay using a Cell Counting Kit-8 (CCK-8; Dojindo) according to the manufacturer's instructions as described previously (47,60).…”

Section: Characteristics Of Ips Cell-derived Nccsmentioning

confidence: 99%

“…Cell phenotypes were identified using flow cytometry as described previously (47,60). Cells were suspended at a density of 1 × 10 6 cells per tube and then incubated with antibodies against the following antigens: cluster of differentiation (CD) 44-hyaluronate receptor, phagocytic glycoprotein-1-(CD44-FITC; BD Biosciences, San Jose, CA, USA), CD73/Ecto-5-prime-nucleotidase (NT5E) (CD73-PE; Biolegend, San Diego, CA, USA), CD90/Thy-1 (CD90-APC; BD Biosciences), CD105/endoglin (CD105-FITC; Biolegend), CD146 -the cell surface glycoprotein MUC18, the melanoma cell adhesion molecule (MCAM)-(CD146-PE; BD Biosciences), the neurotrophin receptor P75 (P75) -CD271, nerve growth factor receptor (NGFR) -(P75-APC; Biolegend), stage-specific embryonic antigen (SSEA)-4 (SSEA-4-Alexa Fluor 647; BD Biosciences), and human natural killer (HNK)-1-the neural cell adhesion molecule (NCAM), CD57-(Sigma-Aldrich).…”

Section: Induction Of Nccs From Ips Cellsmentioning

confidence: 99%

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Induction of neural crest cells from human dental pulp-derived induced pluripotent stem cells

et al. 2017

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We previously generated induced pluripotent stem (iPS) cells from human dental pulp cells of deciduous teeth. Neural crest cells (NCCs) play a vital role in the development of the oral and maxillofacial region. Therefore, NCCs represent a cell source for bone, cartilage, and tooth-related tissue engineering. In this study, we examined whether iPS cells are capable of differentiating into NCCs through modification of the human embryonic stem cell protocol. First, iPS cells were dissociated into single cells and then reaggregated in low-cell-adhesion plates with neural induction medium for 8 days in suspension culture to form neurospheres. The neurospheres were transferred to fibronectin-coated dishes and formed rosette structures. The migrated cells from the rosettes abundantly expressed NCC markers, as evidenced by real-time polymerase chain reaction, immunofluorescence, and flow cytometric analysis. Furthermore, the migrated cells exhibited the ability to differentiate into neural crest lineage cells in vitro. They also exhibited tissue-forming potential in vivo, differentiating into bone and cartilage. Collectively, the migrated cells had similar characteristics to those of NCCs. These results suggest that human dental pulp cell-derived iPS cells are capable of differentiating into NCCs. Therefore, iPS cell-derived NCCs represent cell sources for bone and cartilage tissue engineering.

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Section: Characteristics Of Ips Cell-derived Nccsmentioning

confidence: 99%

Section: Induction Of Nccs From Ips Cellsmentioning

confidence: 99%

Induction of neural crest cells from human dental pulp-derived induced pluripotent stem cells

et al. 2017

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“…We also demonstrated differences in CD105 expression patterns (surface marker) between crown-derived (crown cells) and root-derived dental pulp cells (root cells) of supernumerary teeth. However, there was no difference in CD105 expression patterns between crown cells and root cells in permanent teeth (27), and there is no reported comparable study of crown cells and root cells in primary teeth. We therefore obtained primary teeth that had not undergone root resorption, and separated them into crown and root portions for pulp cell isolation and comparison of pulp cell characteristics based on colony-forming unit-fibroblast (CFU-F) formation, growth potential, expression of surface epitopes, and osteogenic or adipogenic potential.…”

Section: Rna Purification and Analysis By Reverse Transcription Polymmentioning

confidence: 93%

Characterization of mesenchymal progenitor cells in the crown and root pulp of primary teeth

et al. 2015

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The existence of progenitor/mesenchymal stem cells (MSCs) was demonstrated previously in human primary/deciduous teeth. In this study, we examined dental pulp cells from root portion (root cells) of primary teeth without discernible root resorption and compared them with pulp cells from the crown portion (crown cells). Root cells and crown cells were characterized and compared to each other based on progenitor/MSC characteristics and on their generation efficiency of induced pluripotent stem (iPS) cells. Root cells and crown cells included cells manifesting typical progenitor/MSC properties such as osteogenic and adipogenic differentiation potential and clonogenicity. Interestingly, root cells showed a higher expression level of embryonic stem cell marker, KLF4, than crown cells. Moreover, the number of colony-forming unit-fibroblast and cell proliferation rate were higher for root cells than crown cells, and the efficiency of generating iPS cells from root cells was approximately four times higher than that from crown cells. Taken together, these results suggest that root cells from primary teeth show the MSC-like properties and thus could be a potent alternative source for iPS cell generation and the subsequent transplantation therapy.

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“…Cells at passage 2 were seeded onto 6-well plates (1,000 cells per well) and counted using a Cell Counting Kit-8 (WST-8; Dojindo Laboratories, Kumamoto, Japan) according to the manufacturer's instructions on the indicated days as previously described (28). Each test was performed thrice.…”

Section: Cell Proliferation Assaymentioning

confidence: 99%

“…Cell cycle analysis was performed using Click-iT EdU Flow Cytometry Assay Kit (Molecular Probes, Eugene, OR, USA) according to the manufacturer's instructions as previously described (28,29). Briefly, cells at passage 2 were cultured onto 100-mm culture dishes (1.0 × 10 5 cells/dish) in GM for 3 days and treated with 10 mM of EdU.…”

Section: Cell Cycle Analysismentioning

confidence: 99%

Effect of collagenase concentration on the isolation of small adipocytes from human buccal fat pad

et al. 2018

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Dedifferentiated fat (DFAT) cells were isolated from mature adipocytes using the ceiling culture method. Recently, we successfully isolated DFAT cells from adipocytes with a relatively small size (<40 μm). DFAT cells have a higher osteogenic potential than that of medium adipocytes. Therefore, the objective of this study was to determine the optimal concentration of collagenase solution for isolating small adipocytes from human buccal fat pads (BFPs). Four concentrations of collagenase solution (0.01%, 0.02%, 0.1%, and 0.5%) were used, and their effectiveness was assessed by the number of small adipocytes and DFAT cells isolated. The total number of floating adipocytes that dissociated with 0.02% collagenase was 2.5 times of that dissociated with 0.1% collagenase. The number of floating adipocytes with a diameter of ≤29 μm that dissociated with 0.02% collagenase was thrice of those dissociated with 0.1% and 0.5% collagenase. The number of DFAT cells that dissociated with 0.02% collagenase was 1.5 times of that dissociated with 0.1% collagenase. In addition, DFAT cells that dissociated with 0.02% collagenase had a higher osteogenic differentiation potential than those that dissociated with 0.1% collagenase. These results suggest that 0.02% is the optimal collagenase concentration for isolating small adipocytes from BFPs.

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Characterization of mesenchymal progenitor cells in crown and root pulp from human mesiodentes

Abstract: These results indicated that proportion of CD105(+) cells is an effective means of characterizing DP-derived cells in mesiodentes.

Cited by 11 publications

References 38 publications

<b>Induction of neural crest cells from human dental pulp-derived induced pluripotent stem </b><b>cells </b>

<b>Induction of neural crest cells from human dental pulp-derived induced pluripotent stem </b><b>cells </b>

<b>Characterization of mesenchymal progenitor cells in the crown and root pulp of primary </b><b>teeth </b>

Effect of collagenase concentration on the isolation of small adipocytes from human buccal fat pad

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