Isolation and Characterization of Neural Crest-Derived Stem Cells from Dental Pulp of Neonatal Mice
Publication Date
November 08, 2011
Journal
PLOS ONE
Authors
Kajohnkiart Janebodin, Orapin V. Horst, Nicholas Ieronimakis, Gayathri Balasundaram, et al
Volume
6
Issue
11
Pages
e27526
DOI
https://dx.plos.org/10.1371/journal.pone.0027526
Publisher URL
http://journals.plos.org/plosone/article?id=10.1371%2Fjournal.pone.0027526
PubMed
http://www.ncbi.nlm.nih.gov/pubmed/22087335
PubMed Central
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3210810
Europe PMC
http://europepmc.org/abstract/MED/22087335
Web of Science
000297349700057
Scopus
80555129675
Mendeley
http://www.mendeley.com/research/isolation-characterization-neural-crestderived-stem-cells-dental-pulp-neonatal-mice-5
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Mendeley | Further Information

{"title"=>"Isolation and characterization of neural crest-derived stem cells from dental pulp of neonatal mice", "type"=>"journal", "authors"=>[{"first_name"=>"Kajohnkiart", "last_name"=>"Janebodin", "scopus_author_id"=>"54410560100"}, {"first_name"=>"Orapin V.", "last_name"=>"Horst", "scopus_author_id"=>"26635310900"}, {"first_name"=>"Nicholas", "last_name"=>"Ieronimakis", "scopus_author_id"=>"24438010600"}, {"first_name"=>"Gayathri", "last_name"=>"Balasundaram", "scopus_author_id"=>"24437736700"}, {"first_name"=>"Kanit", "last_name"=>"Reesukumal", "scopus_author_id"=>"54380550000"}, {"first_name"=>"Busadee", "last_name"=>"Pratumvinit", "scopus_author_id"=>"35722893000"}, {"first_name"=>"Morayma", "last_name"=>"Reyes", "scopus_author_id"=>"7202844358"}], "year"=>2011, "source"=>"PLoS ONE", "identifiers"=>{"pui"=>"362882598", "issn"=>"19326203", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "doi"=>"10.1371/journal.pone.0027526", "scopus"=>"2-s2.0-80555129675", "pmid"=>"22087335", "sgr"=>"80555129675"}, "id"=>"a6133e38-f41d-3308-a0d4-63fbc51969dc", "abstract"=>"Dental pulp stem cells (DPSCs) are shown to reside within the tooth and play an important role in dentin regeneration. DPSCs were first isolated and characterized from human teeth and most studies have focused on using this adult stem cell for clinical applications. However, mouse DPSCs have not been well characterized and their origin(s) have not yet been elucidated. Herein we examined if murine DPSCs are neural crest derived and determined their in vitro and in vivo capacity. DPSCs from neonatal murine tooth pulp expressed embryonic stem cell and neural crest related genes, but lacked expression of mesodermal genes. Cells isolated from the Wnt1-Cre/R26R-LacZ model, a reporter of neural crest-derived tissues, indicated that DPSCs were Wnt1-marked and therefore of neural crest origin. Clonal DPSCs showed multi-differentiation in neural crest lineage for odontoblasts, chondrocytes, adipocytes, neurons, and smooth muscles. Following in vivo subcutaneous transplantation with hydroxyapatite/tricalcium phosphate, based on tissue/cell morphology and specific antibody staining, the clones differentiated into odontoblast-like cells and produced dentin-like structure. Conversely, bone marrow stromal cells (BMSCs) gave rise to osteoblast-like cells and generated bone-like structure. Interestingly, the capillary distribution in the DPSC transplants showed close proximity to odontoblasts whereas in the BMSC transplants bone condensations were distant to capillaries resembling dentinogenesis in the former vs. osteogenesis in the latter. Thus we demonstrate the existence of neural crest-derived DPSCs with differentiation capacity into cranial mesenchymal tissues and other neural crest-derived tissues. In turn, DPSCs hold promise as a source for regenerating cranial mesenchyme and other neural crest derived tissues.", "link"=>"http://www.mendeley.com/research/isolation-characterization-neural-crestderived-stem-cells-dental-pulp-neonatal-mice-5", "reader_count"=>66, "reader_count_by_academic_status"=>{"Unspecified"=>4, "Professor > Associate Professor"=>4, "Researcher"=>11, "Student > Doctoral Student"=>3, "Student > Ph. D. Student"=>18, "Student > Postgraduate"=>5, "Student > Master"=>14, "Student > Bachelor"=>5, "Professor"=>2}, "reader_count_by_user_role"=>{"Unspecified"=>4, "Professor > Associate Professor"=>4, "Researcher"=>11, "Student > Doctoral Student"=>3, "Student > Ph. D. Student"=>18, "Student > Postgraduate"=>5, "Student > Master"=>14, "Student > Bachelor"=>5, "Professor"=>2}, "reader_count_by_subject_area"=>{"Engineering"=>2, "Unspecified"=>4, "Biochemistry, Genetics and Molecular Biology"=>9, "Materials Science"=>3, "Agricultural and Biological Sciences"=>37, "Medicine and Dentistry"=>10, "Neuroscience"=>1}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>2}, "Materials Science"=>{"Materials Science"=>3}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>10}, "Neuroscience"=>{"Neuroscience"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>37}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>9}, "Unspecified"=>{"Unspecified"=>4}}, "reader_count_by_country"=>{"Colombia"=>1, "United States"=>1, "United Kingdom"=>1, "France"=>1, "India"=>1}, "group_count"=>5}

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Scopus | Further Information

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  • {"files"=>["https://ndownloader.figshare.com/files/715098"], "description"=>"<p>(A–D) After 21 days in osteogenic media, differentiated DPSCs were positive for BSP and OPN in a membranous pattern. (F) Cultures in adipogenic media showed lipid droplets-containing cells positive for Oil Red O. (H and J) Cultures in chondrogenic media showed cells in clusters positive for toluidine blue and COL II. (K) Mineralized nodules were observed in cells treated with the chondrogenic media after 21 days. The inset depicts a mineralized nodule in high magnification. (L) RT-PCR confirmed the staining results. For osteogenic differentiation (Osteogenic D21), RT-PCR showed expression of osteoblast- associated genes <i>Runx2</i>, <i>Osx</i>, <i>Opn</i>, <i>Bsp,</i> and <i>Dmp1</i>. For adipogenic differentiation, induced cells (Adipogenic D21) expressed <i>Pparg2</i> and <i>CEBPa</i>, adipogenic transcription factors, as well as <i>Leptin</i> and <i>Adipsin,</i> markers of adipocytes. For chondrogenic differentiation, treated cells (Chondrogenic D21) expressed chondrocyte-associated genes <i>Sox9</i> and <i>Col2a1</i>. (A, C, E, G, and I) Confluent undifferentiated cells cultured in stem cell media at day 21 (UD D21) expressed some of differentiation genes, but were negative by staining for all differentiation markers and Oil Red O. Cells were counterstained with hematoxylin. RNA isolated from mouse calvarial bone, adipose tissue, and femur was used as positive control for gene expression. Scale bars indicate 100 µm.</p>", "links"=>[], "tags"=>["cultures", "differentiate", "neural", "crest-derived", "mesenchymal"], "article_id"=>385453, "categories"=>["Developmental Biology"], "users"=>["Kajohnkiart Janebodin", "Orapin V. Horst", "Nicholas Ieronimakis", "Gayathri Balasundaram", "Kanit Reesukumal", "Busadee Pratumvinit", "Morayma Reyes"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0027526.g002", "stats"=>{"downloads"=>0, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_DPSC_cultures_differentiate_into_neural_crest_derived_mesenchymal_lineages_/385453", "title"=>"DPSC cultures differentiate into neural crest-derived mesenchymal lineages.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-11-08 01:30:53"}
  • {"files"=>["https://ndownloader.figshare.com/files/362875", "https://ndownloader.figshare.com/files/362913", "https://ndownloader.figshare.com/files/362945", "https://ndownloader.figshare.com/files/362973", "https://ndownloader.figshare.com/files/363008", "https://ndownloader.figshare.com/files/363040", "https://ndownloader.figshare.com/files/363071", "https://ndownloader.figshare.com/files/363126", "https://ndownloader.figshare.com/files/363178", "https://ndownloader.figshare.com/files/363210", "https://ndownloader.figshare.com/files/363256", "https://ndownloader.figshare.com/files/363299", "https://ndownloader.figshare.com/files/363352", "https://ndownloader.figshare.com/files/363404", "https://ndownloader.figshare.com/files/363452"], "description"=>"<div><p>Dental pulp stem cells (DPSCs) are shown to reside within the tooth and play an important role in dentin regeneration. DPSCs were first isolated and characterized from human teeth and most studies have focused on using this adult stem cell for clinical applications. However, mouse DPSCs have not been well characterized and their origin(s) have not yet been elucidated. Herein we examined if murine DPSCs are neural crest derived and determined their in vitro and in vivo capacity. DPSCs from neonatal murine tooth pulp expressed embryonic stem cell and neural crest related genes, but lacked expression of mesodermal genes. Cells isolated from the <em>Wnt1-Cre/R26R-LacZ</em> model, a reporter of neural crest-derived tissues, indicated that DPSCs were <em>Wnt1</em>-marked and therefore of neural crest origin. Clonal DPSCs showed multi-differentiation in neural crest lineage for odontoblasts, chondrocytes, adipocytes, neurons, and smooth muscles. Following in vivo subcutaneous transplantation with hydroxyapatite/tricalcium phosphate, based on tissue/cell morphology and specific antibody staining, the clones differentiated into odontoblast-like cells and produced dentin-like structure. Conversely, bone marrow stromal cells (BMSCs) gave rise to osteoblast-like cells and generated bone-like structure. Interestingly, the capillary distribution in the DPSC transplants showed close proximity to odontoblasts whereas in the BMSC transplants bone condensations were distant to capillaries resembling dentinogenesis in the former vs. osteogenesis in the latter. Thus we demonstrate the existence of neural crest-derived DPSCs with differentiation capacity into cranial mesenchymal tissues and other neural crest-derived tissues. In turn, DPSCs hold promise as a source for regenerating cranial mesenchyme and other neural crest derived tissues.</p> </div>", "links"=>[], "tags"=>["characterization", "neural", "crest-derived", "cells", "dental", "pulp", "neonatal", "mice"], "article_id"=>131710, "categories"=>["Developmental Biology"], "users"=>["Kajohnkiart Janebodin", "Orapin V. Horst", "Nicholas Ieronimakis", "Gayathri Balasundaram", "Kanit Reesukumal", "Busadee Pratumvinit", "Morayma Reyes"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0027526.s001", "https://dx.doi.org/10.1371/journal.pone.0027526.s002", "https://dx.doi.org/10.1371/journal.pone.0027526.s003", "https://dx.doi.org/10.1371/journal.pone.0027526.s004", "https://dx.doi.org/10.1371/journal.pone.0027526.s005", "https://dx.doi.org/10.1371/journal.pone.0027526.s006", "https://dx.doi.org/10.1371/journal.pone.0027526.s007", "https://dx.doi.org/10.1371/journal.pone.0027526.s008", "https://dx.doi.org/10.1371/journal.pone.0027526.s009", "https://dx.doi.org/10.1371/journal.pone.0027526.s010", "https://dx.doi.org/10.1371/journal.pone.0027526.s011", "https://dx.doi.org/10.1371/journal.pone.0027526.s012", "https://dx.doi.org/10.1371/journal.pone.0027526.s013", "https://dx.doi.org/10.1371/journal.pone.0027526.s014", "https://dx.doi.org/10.1371/journal.pone.0027526.s015"], "stats"=>{"downloads"=>8, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Isolation_and_Characterization_of_Neural_Crest_Derived_Stem_Cells_from_Dental_Pulp_of_Neonatal_Mice/131710", "title"=>"Isolation and Characterization of Neural Crest-Derived Stem Cells from Dental Pulp of Neonatal Mice", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2011-11-08 00:28:30"}
  • {"files"=>["https://ndownloader.figshare.com/files/715351"], "description"=>"<p>Treatment of DPSC clones with osteogenic media resulted in different outcomes compared to the differentiation of non-clonal DPSCs. (A–D) Clones did not produce positive cells for BSP and OPN while non-clonal populations showed clusters of positive cells for these markers. (E–J) Non-clonal and all clonal DPSC cultures in osteogenic media stained positively for DMP1, DSP, and OCN. (K) Corresponding to staining results, RT-PCR shows the expression of differentiation genes following treatment of clones with osteogenic media, lack of <i>Bsp</i>, but strong expression of <i>Dmp1</i> was observed in the differentiation of DPSC clones (Osteogenic D21). (L and M) Non-clonal and 4 out of 5 clones (C5–C8) stained positively for COL II after treatment with chondrogenic media. (N) RT-PCR shows strong expression of chondrogenic markers, <i>Sox9</i> and <i>Col2a1</i>, in DPSCs cultured in chondrogenic media (Chondrogenic D21). Staining and RT-PCR are all derived from a representative clone (C6). RNA isolated from mouse mandible and femur was used as positive control for gene expression. Scale bars indicate 100 µm.</p>", "links"=>[], "tags"=>["clonal", "dpsc", "cultures", "differentiate", "mesenchymal"], "article_id"=>385708, "categories"=>["Developmental Biology"], "users"=>["Kajohnkiart Janebodin", "Orapin V. Horst", "Nicholas Ieronimakis", "Gayathri Balasundaram", "Kanit Reesukumal", "Busadee Pratumvinit", "Morayma Reyes"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0027526.g004", "stats"=>{"downloads"=>0, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Non_clonal_and_clonal_DPSC_cultures_differentiate_into_mesenchymal_lineages_/385708", "title"=>"Non-clonal and clonal DPSC cultures differentiate into mesenchymal lineages.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-11-08 01:35:08"}
  • {"files"=>["https://ndownloader.figshare.com/files/716564"], "description"=>"<p>(A–D) Neuronal-induced cells stained positively for neuronal markers; neurofilament and S100 (in red). (E) The neurofilament staining was confirmed by the expression of neurofilament-light (NFL) and -heavy (NFH) of treated cells in neurogenic media after 21 days (N D21) (F–I) DPSCs in smooth muscle differentiation media showed smooth muscle-like phenotype that stained positively in red fluorescence for smooth muscle actin, smooth muscle myosin heavy chain, calponin, caldesmon. DAPI used for nuclei staining is depicted in blue. (J) Corresponding to immunofluorescence, Q-RT-PCR revealed the up-regulation of smooth muscle-specific genes, <i>SRF, Sm22-alpha, Sma, SMHC,</i> and <i>calponin</i> in DPSCs after cultured in smooth muscle media. Scale bars indicate 100 µm. RQ values were normalized by the expression of mouse smooth muscle cells. <i>GAPDH</i> was used for the internal control. Error bars represent ±SEM.</p>", "links"=>[], "tags"=>["dpscs", "gave", "neural", "crest-derived", "non-mesenchymal"], "article_id"=>386919, "categories"=>["Developmental Biology"], "users"=>["Kajohnkiart Janebodin", "Orapin V. Horst", "Nicholas Ieronimakis", "Gayathri Balasundaram", "Kanit Reesukumal", "Busadee Pratumvinit", "Morayma Reyes"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0027526.g012", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Wnt1_marked_DPSCs_gave_rise_into_neural_crest_derived_non_mesenchymal_lineages_/386919", "title"=>"<i>Wnt1</i>-marked DPSCs gave rise into neural crest-derived non-mesenchymal lineages.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-11-08 01:55:19"}
  • {"files"=>["https://ndownloader.figshare.com/files/716311"], "description"=>"<p>(A) Immunofluorescence showed <i>Wnt1-Cre/R26R-LacZ</i> derived dental pulp cells and odontoblasts expressing β-galactosidase (B-gal, in green). The co-expression of β-galactosidase and mesenchymal stem cell marker, CD44, in plasma membrane (in red) was observed in DPSCs located in sub-odontoblastic (arrowheads) and perivascular regions (arrows). (B) In <i>Tie2-GFP</i> dental pulp only vessels express green fluorescent protein (GFP), CD44 positive cells were particularly located in sub-odontoblastic (arrowheads) and perivascular areas (arrows). (C and D) The co-localization of CD44 (in red) and MSI1 (in pseudo-color cyan), a neural crest-related marker, was observed in cells located in sub-odontoblastic (arrowheads) and perivascular areas (arrows) of both <i>Wnt1-Cre/R26R-LacZ</i> and <i>Tie2-GFP</i>, respectively. (E) Staining with IgG isotypes are shown as negative control. (F) <i>Tie2-GFP</i> derived dental pulp cells did not express β-galactosidase, but blood vessels in this transgenic mouse were labeled by GFP (arrows). <i>D</i>  =  Dentin, <i>DP</i>  =  Dental pulp, <i>OD</i>  =  Odontoblasts. Scale bars indicate 100 µm.</p>", "links"=>[], "tags"=>["neural", "crest-derived", "cells", "dental", "pulp", "neonatal"], "article_id"=>386664, "categories"=>["Developmental Biology"], "users"=>["Kajohnkiart Janebodin", "Orapin V. Horst", "Nicholas Ieronimakis", "Gayathri Balasundaram", "Kanit Reesukumal", "Busadee Pratumvinit", "Morayma Reyes"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0027526.g010", "stats"=>{"downloads"=>0, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Localization_of_neural_crest_derived_stem_cells_in_dental_pulp_of_neonatal_mice_/386664", "title"=>"Localization of neural crest-derived stem cells in dental pulp of neonatal mice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-11-08 01:51:04"}
  • {"files"=>["https://ndownloader.figshare.com/files/715490"], "description"=>"<p>(B) All clones showed smooth muscle-like cells positive for smooth muscle actin (in red). (D, F, and H) Three clones (C6–C8) showed positive cells for neurofilament (NF-160/200), S100, and NG2. (I) The NF staining was confirmed by RT-PCR of neurofilament-light (NFL) and -heavy (NFH). (A, C, E, and G) Undifferentiated clones were negative for smooth muscle and neuronal markers. DAPI used for nuclei staining is depicted in blue. Scale bars indicate 100 µm.</p>", "links"=>[], "tags"=>["clones", "differentiate", "neural", "crest", "non-mesenchymal"], "article_id"=>385845, "categories"=>["Developmental Biology"], "users"=>["Kajohnkiart Janebodin", "Orapin V. Horst", "Nicholas Ieronimakis", "Gayathri Balasundaram", "Kanit Reesukumal", "Busadee Pratumvinit", "Morayma Reyes"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0027526.g005", "stats"=>{"downloads"=>0, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_DPSC_clones_differentiate_into_neural_crest_non_mesenchymal_lineages_/385845", "title"=>"DPSC clones differentiate into neural crest non-mesenchymal lineages.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-11-08 01:37:25"}
  • {"files"=>["https://ndownloader.figshare.com/files/716176"], "description"=>"<p>(A) DPSCs first isolated from 4–10 day-old <i>Wnt1-Cre/R26R-LacZ</i> showed a majority of <i>Wnt1</i>-marked cells in primary culture on day 8 (d8) which stained positively for β-galactosidase with X-gal. A significant minority (10%) of negative cells was also observed in early cultures (arrowheads). (B) Bone marrow stromal cells (BMSCs) isolated from the same <i>Wnt1-Cre/R26R-LacZ</i> cultured in the same condition for 8 days showed negative staining for X-gal. DPSCs were successfully cloned on day 7–10. Following several passages until day 40 (d40), all clones generated in this experiment (n = 10) were 100% positive for X-gal. (C and D) Two different <i>Wnt1</i> marked clones were shown. (E) A representative DPSC clone from C57BL6 was negative for X-gal. (F) RT-PCR demonstrated that DPSC clones strongly expressed most of neural crest-related genes, <i>Msi1, Sox10, TrkC, Twist, Snail,</i> and slightly expressed <i>LNGFR</i>, but not in BMSCs. Both DPSCs and BMSCs expressed <i>CD105</i>, a known mesenchymal stem cell marker. DPSC clones also expressed pluripotent stem cell genes, <i>Nanog, Klf4, Sox2</i> and <i>c-Myc.</i> (G–J) Immunofluorescence showed positive staining of mesenchymal stem cell markers, CD44, CD73, CD105, and neural crest-related marker, MSI1 in DPSC clones (in red). (K) Staining with IgG isotype was used as negative control. DAPI used for nuclei staining is depicted in blue. Scale bars indicate 50 µm.</p>", "links"=>[], "tags"=>["clones"], "article_id"=>386534, "categories"=>["Developmental Biology"], "users"=>["Kajohnkiart Janebodin", "Orapin V. Horst", "Nicholas Ieronimakis", "Gayathri Balasundaram", "Kanit Reesukumal", "Busadee Pratumvinit", "Morayma Reyes"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0027526.g009", "stats"=>{"downloads"=>0, "page_views"=>13, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_DPSC_clones_from_Wnt1_Cre_R26R_LacZ_mice_/386534", "title"=>"DPSC clones from <i>Wnt1-Cre/R26R-LacZ</i> mice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-11-08 01:48:54"}
  • {"files"=>["https://ndownloader.figshare.com/files/715796"], "description"=>"<p>Panel A shows the staining of 5-week subcutaneous (SC) clonal DPSC transplanted tissues. (A–C) Three DPSC clones (C6–C8) secreted extracellular matrices which were positive for DMP1, slightly positive for DSP and BSP by immunoperoxidase staining. (D–F) High magnification images correspond to the rectangular areas of A–C. (D) Strong DMP1 staining was seen in the matrices. (E and F, arrows) Strong positive staining for DSP and BSP were observed in the transplanted cells. Panel B shows the staining of 12-week SC transplantations of non-clonal DPSCs, clonal DPSCs, and BMSCs with HAp/TCP. (G–I) Only HAp/TCP transplantation without donor cells did not show any extracellular matrices positive for dentin and bone proteins. (J–O) Both non-clonal and clonal DPSCs secreted extracellular matrices which were strongly positive, but different intensity, to DMP1, DSP, and BSP. All insets in each figure show high magnification images of the transplanted tissues to illustrate the morphology and positive intracellular staining of transplanted cells. (K, M, and N insets) transplanted cells were elongated, polarized, and showed very close relation to scaffold surface, resembling odontoblast-like cell morphology. (K, L, M, N, and O insets) Positive staining of DMP1, DSP, and BSP was seen in transplanted DPSCs. Some cells showed elongation and polarization. (P, arrowheads and arrows) BMSC transplanted cells showed osteoblast-like cells (arrowheads) lining bone surface, and osteocyte-like cells (arrows) in lacunae which stained positively for DMP1. (Q) Negative staining for DSP was observed in bone matrix and transplanted BMSC cells. (R, inset) Positive staining for BSP was observed in bone matrix secreted by transplanted BMSCs. (S–V) Transplanted sections stained with only anti-rabbit biotinylated antibody or IgG isotype were used as negative control. <i>HAp/TCP</i>  =  Hydroxyapatite/tricalcium phosphate. Scale bars indicate 200 µm.</p>", "links"=>[], "tags"=>["subcutaneous", "dpsc", "bmsc", "transplanted"], "article_id"=>386151, "categories"=>["Developmental Biology"], "users"=>["Kajohnkiart Janebodin", "Orapin V. Horst", "Nicholas Ieronimakis", "Gayathri Balasundaram", "Kanit Reesukumal", "Busadee Pratumvinit", "Morayma Reyes"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0027526.g007", "stats"=>{"downloads"=>1, "page_views"=>14, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Immunohistochemistry_of_subcutaneous_DPSC_and_BMSC_transplanted_tissues_/386151", "title"=>"Immunohistochemistry of subcutaneous DPSC and BMSC transplanted tissues.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-11-08 01:42:31"}
  • {"files"=>["https://ndownloader.figshare.com/files/716036"], "description"=>"<p>The average distance of donor nuclei in condensed matrices to nearest capillaries was measured using ImageJ v1.43u (Wayne Rasband, NIH; <a href=\"http://rsb.info.nih.gov/ij\" target=\"_blank\">http://rsb.info.nih.gov/ij</a>). The distances between donor nuclei to blood vessels were represented by yellow lines. (A and B) In the 12-week DPSC transplants, abundant microvessels with fenestrated morphology were seen in close proximity to odontoblast-like cells in both non clonal and clonal DPSC transplants. (C) In BMSC transplants microvessels were predominantly found in areas rich in adipocytes whereas mineralized areas formed by BMSCs were distant to microvessels. Microvessels were mature as circulating red blood cells can be seen throughout in both DPSC and BMSC transplants. (D) Abundant microvessels with fenestrated morphology containing circulating red blood cells (insets) are seen in close proximity to mineralized matrix formed by DPSCs. (E) The bar graph shows that the average distance from odontoblast nuclei to nearest capillary was significantly close (9 µm ± 4.49) and consistent across all DPSC transplants whereas the average distance of osteocyte nuclei in bone condensations to nearest microvessels were significantly farther (187 µm ± 76.88). The average distances from donor cell nuclei to blood vessels in non-clonal DPSCs (n = 5, n; numbers of measured area) or clonal DPSCs (n = 7) and BMSCs (n = 5) are statistically significant. Student's t-test calculated * P≤0.05. Error bars represent ±SEM. Scale bars indicate 25 µm.</p>", "links"=>[], "tags"=>["nuclei", "vessels", "dpsc", "bmsc"], "article_id"=>386388, "categories"=>["Developmental Biology"], "users"=>["Kajohnkiart Janebodin", "Orapin V. Horst", "Nicholas Ieronimakis", "Gayathri Balasundaram", "Kanit Reesukumal", "Busadee Pratumvinit", "Morayma Reyes"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0027526.g008", "stats"=>{"downloads"=>0, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Proximity_of_donor_cell_nuclei_to_blood_vessels_in_DPSC_and_BMSC_transplantation_/386388", "title"=>"Proximity of donor cell nuclei to blood vessels in DPSC and BMSC transplantation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-11-08 01:46:28"}
  • {"files"=>["https://ndownloader.figshare.com/files/714998"], "description"=>"<p>Gene expression of fresh dental pulp and DPSCs cultured several passages from three isolations (#1, #2 and #3) are shown. (A) Stem cell genes, <i>Oct4</i>, <i>Sox2</i>, <i>Klf4</i>, <i>Nanog</i> and <i>c-Kit,</i> were observed; however, <i>Oct4</i> was inconsistent and transient from early to late passages. (B) In contrast to neural crest developmental genes <i>Gsc</i> and G<i>ATA6,</i> early mesodermal developmental genes <i>Brachyury</i> and <i>Mesp2</i> were not expressed. (C) Neural crest-related genes <i>TrkC, Pdgfra, LNGFR, Twist, Slug, Snail, Msi1</i>, and <i>NCAM</i>, were continuously expressed in all three non-clonal DPSCs. RNA isolated from mouse embryonic stem cells (mESCs) and salivary gland were used as positive control for the expression of stem cell/early developmental genes and neural crest genes, respectively. The PCR reaction without template was used as negative control.</p>", "links"=>[], "tags"=>["dpsc"], "article_id"=>385357, "categories"=>["Developmental Biology"], "users"=>["Kajohnkiart Janebodin", "Orapin V. Horst", "Nicholas Ieronimakis", "Gayathri Balasundaram", "Kanit Reesukumal", "Busadee Pratumvinit", "Morayma Reyes"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0027526.g001", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Gene_profile_of_DPSC_cultures_/385357", "title"=>"Gene profile of DPSC cultures.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-11-08 01:29:17"}
  • {"files"=>["https://ndownloader.figshare.com/files/715630"], "description"=>"<p>Non-clonal and clonal DPSCs (C6–C8) were labeled with PKH-26 before transplantation to track cells <i>in vivo</i>. Panel A shows 5-week subcutaneous (SC) transplantation of DPSC clones with HAp/TCP. (A and B) PKH-26+ cells SC transplanted with HAp/TCP were identified in bright red fluorescent color whereas the HAp/TCP scaffold without cells was devoid of red fluorescent cells despite some autofluoresence derived from HAp/TCP. The HAp/TCP scaffold without cells did not show mineralized tissues (data not shown). Most of transplanted cells surrounded HAp/TCP. (C) These clonal cells generated collagen-forming matrix shown in blue of Masson's trichrome. (D, white arrowheads) Polarized light was used to confirm collagen formation. Panel B shows 12-week SC transplantation of non-clonal DPSCs, clonal DPSCs, and BMSCs with HAp/TCP. (E–G) H&E staining showed the morphology of tissues created by non-clonal, clonal DPSCs and BMSCs. Capillaries and small blood vessels near mineralized matrices were found in DPSC transplants. In contrast, BMSCs formed bone condensations near HAp/TCP devoid of vessels. In BMSC transplants capillaries were found mainly in the adipose tissue. The black arrowheads indicate the location of capillaries and small blood vessels. (H–J) Masson's trichrome showed the intensive staining of collagenous extracellular matrices generated by both DPSCs and BMSCs. (K–M) polarized light images with high magnification of transplanted tissues showed different collagen arrangement produced by DPSCs and BMSCs. (K and L, white arrowheads, and M) The white arrowheads indicate the direction of polarized collagen fibers created by DPSCs run perpendicularly to HAp/TCP while that created by BMSCs do not run perpendicularly to scaffold surface. DAPI used for nuclei staining is depicted in blue. <i>HAp/TCP</i>  =  Hydroxyapatite/tricalcium phosphate. Scale bars indicate 200 µm.</p>", "links"=>[], "tags"=>["transplantation", "dpscs"], "article_id"=>385986, "categories"=>["Developmental Biology"], "users"=>["Kajohnkiart Janebodin", "Orapin V. Horst", "Nicholas Ieronimakis", "Gayathri Balasundaram", "Kanit Reesukumal", "Busadee Pratumvinit", "Morayma Reyes"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0027526.g006", "stats"=>{"downloads"=>0, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Subcutaneous_transplantation_of_DPSCs_and_BMSCs_/385986", "title"=>"Subcutaneous transplantation of DPSCs and BMSCs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-11-08 01:39:46"}
  • {"files"=>["https://ndownloader.figshare.com/files/715221"], "description"=>"<p>(B and D) Neuronal-induced cells stained positively for neuronal markers; N-CAM ( in green), and GABA (in red). (F) DPSCs in smooth muscle differentiation media showed smooth muscle-like phenotype that stained positively for smooth muscle actin (in red). (A, C, and E) Undifferentiated cells in stem cell media at day 21 did not stain positively for any of the three differentiation markers. DAPI used for nuclei staining is depicted in blue. Scale bars indicate 100 µm.</p>", "links"=>[], "tags"=>["cultures", "differentiate", "neural", "crest", "non-mesenchymal"], "article_id"=>385569, "categories"=>["Developmental Biology"], "users"=>["Kajohnkiart Janebodin", "Orapin V. Horst", "Nicholas Ieronimakis", "Gayathri Balasundaram", "Kanit Reesukumal", "Busadee Pratumvinit", "Morayma Reyes"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0027526.g003", "stats"=>{"downloads"=>0, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_DPSC_cultures_differentiate_into_neural_crest_non_mesenchymal_lineages_/385569", "title"=>"DPSC cultures differentiate into neural crest non-mesenchymal lineages.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-11-08 01:32:49"}
  • {"files"=>["https://ndownloader.figshare.com/files/716831"], "description"=>"<p>(A) Base on the <i>Wnt1-Cre/R26R-LacZ</i> mouse model, neural crest gene expression, and differentiation capacity, dental pulp stem cell (DPSC) populations contain a majority of neural crest-derived cells (approximately 90%) (shown in blue) while the remaining cell populations were derived from non-neural crest origin (shown in white). In our condition, only neural crest-derived cells can survive and proliferate. After <i>in vitro</i> differentiation, these cells can give rise to neural crest-lineages, neuronal-like, smooth muscle/pericyte-like, odontoblast-like cells, including classical mesenchymal cell lineages, osteoblast-like, chondrocyte-like, and adipocytes. Nonetheless, in this study the differentiation capacity of non-neural crest-derived mesenchymal stem cell (MSC)-like cells was not characterized. (B) <i>In vivo</i> models show the different capacity between DPSCs and bone marrow stromal cells (BMSCs). Based on the cell/tissue morphology and antibody staining, DPSCs formed dentin-like matrix composed of odontoblast-like cells and pericytes associated with microvessels (close distance about 9–12 µm) recapitulating dentinogenesis. On the other hand, the formation of bone condensation from BMSCs occurred in avascular areas (far distant, about 100–200 µm), resembling early osteogenesis. This model illustrates how two different types of mesenchymal stem cells derived from different origins form different niches and matrices when transplanted under the same conditions. Interestingly, these tissue-specific stem cells recapitulate formation of their own tissue (organogenesis). Understanding the mechanisms of this “memory” of their tissues of origin may be pivotal information for tissue regeneration and cell therapy.</p>", "links"=>[], "tags"=>["models", "neural", "crest-derived", "cells", "murine", "dental", "pulp"], "article_id"=>387184, "categories"=>["Developmental Biology"], "users"=>["Kajohnkiart Janebodin", "Orapin V. Horst", "Nicholas Ieronimakis", "Gayathri Balasundaram", "Kanit Reesukumal", "Busadee Pratumvinit", "Morayma Reyes"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0027526.g014", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Summary_of_results_and_working_models_of_neural_crest_derived_stem_cells_isolated_from_murine_dental_pulp_tissue_/387184", "title"=>"Summary of results and working models of neural crest-derived stem cells isolated from murine dental pulp tissue.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-11-08 01:59:44"}
  • {"files"=>["https://ndownloader.figshare.com/files/716448"], "description"=>"<p>(A) Double staining showed that X-gal+ treated cells in adipogenic media demonstrated lipid droplets-containing cells positive for Oil Red O. (B) Chondrogenic differentiation by monolayer method showed that a cluster of chondrocyte-like cells derived from DPSCs secreted extracellular matrices positive for COLII. (C and D) DPSCs cultured in chondrogenic media by pellet method formed aggregates containing highly proteoglycan which stained positively for toluidine blue and alcian blue, respectively. (E) Differentiated DPSCs in osteo-odontogenic media stained with IgG isotype as negative control showed X-gal positive staining. (F and G) DPSCs treated in osteo-odontogenic media for 21 days secreted dentin matrices stained positively for DMP1 and DSP; those extracellular matrices were not found in cells cultured in stem cell media. (H–J) RT-PCR from each differentiation confirmed the staining results. DPSCs treated in each differentiation media up-regulated specific adipogenic (A D14), chondrogenic (C D21), and osteo-odontogenic (B D21) genes after 14 to 21-day culture. RNA of mouse adipose tissue, femur, and mandible was used as positive control while the reaction without cDNA was used as negative control. Scale bars indicate 100 µm.</p>", "links"=>[], "tags"=>["neural", "crest-derived", "mesenchymal", "lineages"], "article_id"=>386803, "categories"=>["Developmental Biology"], "users"=>["Kajohnkiart Janebodin", "Orapin V. Horst", "Nicholas Ieronimakis", "Gayathri Balasundaram", "Kanit Reesukumal", "Busadee Pratumvinit", "Morayma Reyes"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0027526.g011", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Multi_differentiation_capacity_in_neural_crest_derived_mesenchymal_lineages_of_Wnt1_marked_DPSCs_/386803", "title"=>"Multi-differentiation capacity in neural crest-derived mesenchymal lineages of <i>Wnt1</i>-marked DPSCs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-11-08 01:53:23"}
  • {"files"=>["https://ndownloader.figshare.com/files/716699"], "description"=>"<p>The <i>Wnt1</i>-marked DPSCs were subcutaneously transplanted in HAp/TCP scaffolds and analyzed 5 weeks post-transplantation. (A) anti β-galactosidase staining (in green) indicated the location of transplanted cells in DPSC transplantation. (B and C) Negative staining of anti β-galactosidase were observed in both transplantation of HAp/TCP only and IgG isotype control, respectively. (D and E) Co-localization of cells positive for β-galactosidase (in green) and CD44 (in red), as well as, MSI1 (in green) and CD44 (in red) were shown in odontoblast-like cells surrounding HAp/TCP scaffold surfaces in DPSC transplants. (F–H) H&E staining demonstrated the morphology of transplanted tissues in DPSC transplants. The transplanted DPSCs concentrated near HAp/TCP surfaces, and secreted extracellular matrices. Some of transplanted cells were elongated, polarized, and located perpendicularly to HAp/TCP surfaces, resembling odontoblast-like cells (indicated by arrows). Many loose connective tissue areas containing small blood vessels (indicated by arrowheads) surrounded by transplanted cells were found in DPSC transplants, resembling dental pulp-like tissues. (I) transplantations of HAp/TCP without cells were devoid of dense extracellular matrix. (J–M) Masson's trichrome staining confirmed the formation of collagen matrices found in DPSC transplants, but not in the transplantation of HAp/TCP only, respectively. (J–L) DPSC transplants showed elongated and polarized transplanted cells (indicated by arrows). (N–Q) DMP and DSP staining was strongly positive in DPSC transplants, but negative in the transplantation of HAp/TCP only, respectively. (O) White arrowheads illustrate polarized transplanted cells positive for DSP. (R) Staining with IgG isotype was used as negative control. DAPI used for nuclei staining is depicted in blue. The dot lines represent a boundary between extracellular matrices secreted by DPSCs and dental pulp-like structure. <i>DP</i>  =  Dental pulp-like structure, <i>HAp/TCP</i>  =  Hydroxyapatite/tricalcium phosphate. Scale bars indicate 100 µm.</p>", "links"=>[], "tags"=>["dpscs", "gave", "odontoblast-like", "cells", "generated", "dentin-like"], "article_id"=>387046, "categories"=>["Developmental Biology"], "users"=>["Kajohnkiart Janebodin", "Orapin V. Horst", "Nicholas Ieronimakis", "Gayathri Balasundaram", "Kanit Reesukumal", "Busadee Pratumvinit", "Morayma Reyes"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0027526.g013", "stats"=>{"downloads"=>0, "page_views"=>18, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Wnt1_marked_DPSCs_gave_rise_to_odontoblast_like_cells_and_generated_dentin_like_structure_in_vivo_/387046", "title"=>"<i>Wnt1</i>-marked DPSCs gave rise to odontoblast-like cells and generated dentin-like structure in vivo.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-11-08 01:57:26"}

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