This Article Abstract Full Text Full Text (PDF) Data Supplement Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Add to Saved Citations Download to citation manager Request Permissions Request Reprints Add to My Marked Citations Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Citing Articles via Scopus Google Scholar Articles by Gronthos, S. Articles by Shi, S. Search for Related Content PubMed PubMed Citation Articles by Gronthos, S. Articles by Shi, S. Pubmed/NCBI databases Substance via MeSH Medline Plus Health Information Stem Cells Social Bookmarking                         What's this?

Stem Cell Properties of Human Dental Pulp Stem Cells

¹ Craniofacial and Skeletal Diseases Branch, Building 30, Room 228, and
² CIinical Research Core, NIDCR, NIH, Bethesda, MD 20892, USA;
³ School of Dentistry, University of California at San Francisco, USA;
⁴ Department of Anatomy and Developmental Biology, University College London, UK;

View larger version (165K): [in this window] [in a new window]   Figure 1. Characterization of DPSC transplant. (A) The dentin (D) generated in DSPC transplants was associated with connective tissue (CT), organized in a fashion similar to that of the tissue structure in dental pulp, containing odontoblasts (arrows) lining the surface of dentin (D), fibrous tissue, and blood vessels (triangles). (B) BMSSC transplants showed newly generated bone (B) surrounding hematopoietic marrow elements (HM) containing adipocytes (arrows). (C) DPSC transplants show negative staining for tartrate-resistant acid phosphatase (TRAP). (D) TRAP-positive osteoclasts were found in BMSSC transplants (arrows). (E,F) Back-scattered electron microscopy of DPSC and BMSSC transplants, respectively. A mineralized globular dentin-like structure (D) was found around the surfaces of HA/TCP (HA) (E) and mineralized bone lamellae (B) covered the surfaces of HA/TCP (HA) in BMSSC transplants (F).

View larger version (73K): [in this window] [in a new window]   Figure 2. Isolation of stromal-like cells from DPSC transplants. Cells from DPSC transplants were isolated by fluorescence-activated cell sorting (FACS) as described in MATERIALS & METHODS. The majority of cells (85%) isolated from three-month DPSC transplants reacted with human-specific anti-CD29 monoclonal antibody (A) while the remaining cells (15%) reacted with the mouse-specific anti-CD29 antibody (B) using FACS. Similar results were obtained with human-specific alu (C) and mouse-specific pf1 (D) in situ hybridization, where the majority of cells were of human and far fewer cells of mouse. Trichrome staining (E), human alu in situ hybridization (F), and immunohistochemical staining (G) showed that these stromal-like cells, re-transplanted into immunocompromised mice, differentiated into alu-positive odontoblasts (triangles) that generated dentin (D) and linked to pulp-like connective tissue (CT). The newly generated dentin contained organized collagen fibers running perpendicular to the forming surface (blue color in E) and was positive for human DSP antibody staining (arrows in G).

View larger version (95K): [in this window] [in a new window]   Figure 3. Adipogenic and neural differentiation of human DPSCs. In an adipogenic medium, DPSCs formed Oil red O-positive lipid clusters (arrows in A) and showed a significant up-regulation of PPAR2 and lipoprotein lipase (LPL) in the induced group (ID) as compared with the control group (CT) by RT-PCR (B). No measurable levels of lipoprotein lipase were detected in the control group (B). DPSCs were also immunostained for Nestin (C) and GFAP (D), while BMSSCs were immunoreactive only for Nestin (E) and not for GFAP (F). Similar results were obtained from RT-PCR (G). DPSCs (DP) expressed both Nestin and GFAP at a high level when compared with the level of Nestin and the undetectable level of GFAP in BMSSCs (BM). GAPDH, a housekeeper gene, served as a PCR amplification control. M = markers.

View larger version (62K): [in this window] [in a new window]   Figure 4. Cloning efficiency of DPSCs. (A) Of 15 selected single-colony-derived strains of DPSCs, only 3 were capable of proliferating over 20 population doublings (PD) (strains #C, #E, and #J). These 3 strains proliferated beyond 20 PDs (final PD is not yet identified), but all other strains proliferated only between 10 and 20 PD. These strains were not able to generate enough cells for study of their developmental potential. Altogether, 12 single-colony-derived DPSC strains (including strains #C, #E, and #J presented here), capable of exceeding 20 PD, were developed and tested with respect to their ability to form a dentin-pulp-like complex in vivo. After approximately 25 PDs, they were transplanted subcutaneously into immunocompromised mice for 8 wks. Two-thirds (8 of 12) of these single-colony-derived DPSC strains formed the same amount of dentin as multi-colony DPSCs (B) and 1/3 (4 of 12) generated only a limited amount of dentin (C). Newly formed dentin (arrows) and odontoblasts (triangles) were immunoreactive for DSP antibody in single-colony-derived DPSC transplants (D). Positive control, demineralized human dentin section, showing the peritubular areas immunostained with human DSP antibody (E, arrows).

Journal of Dental Research, Vol. 81, No. 8, 531-535 (2002)
DOI: 10.1177/154405910208100806


CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    Twitter    What's this?