미성숙 매복지치의 치낭, 치수, 치근유두 조직에서 다능성 줄기세포의 분리와 특성화에 대한 연구
Isolation and characterization of human dental tissue-derived stem cells in the impacted wisdom teeth: comparison of dental follicle, dental pulp, and root apical papilla-derived cells
1
조회
0
다운로드
7
인용
0
참고문헌
키워드
초록
Introduction: The first aim of this study was to isolate the dental tissue-derived stem cells from the dental follicle (DF), dental pulp (DP), and root apical papilla (RAP) of the extracted wisdom teeth. Second was to evaluate their characterization with the expressions of transcription factors and cell surface markers. Finally, their ability of the in vitro multi-lineage differentiations into osteogenic and adipogenic cells were compared, respectively. Materials and Methods: Dental tissues, including dental follicle, dental pulp, and root apical papilla, were separated in the extracted wisdom teeth. These three dental tissues were cultured in Dulbecco’s modified Eagle’s medium (DMEM) with supplements, respectively. After passage 3, the homogeneous shaped dental tissue-derived cells were analyzed the expression of transcription factors (Oct-4, Nanog and Sox-2) and cell surface markers (CD44, CD90 and CD105) with reverse transcription polymerase chain reaction (RT-PCR) and fluorescence-activated cell sorting (FACS) analysis. In order to evaluate in vitro multi-lineage differentiations, the culture media were changed to the osteogenic and adipogenic induction mediums when the dental tissue-derived cells reached to passage 3. The characteristics of these three dental tissue-derived cells were compared with immunohistochemistry. Results: During primary culture, heterogenous and colony formatted dental tissue-derived cells were observed in the culture plates. After passage 2or 3, homogenous spindle-like cells were observed in all culture plates. Transcription factors and mesenchymal stem cell markers were positively observed in all three types of dental tissue-derived cells. However, the quantity of expressed transcription factors was most large in RAP-derived cells. In all three types of dental tissue-derived cells, osteogenic and adipogenic differentiations were observed after treatment of specific induction media. In vitro adipogenic differentiation was similar among these three types of cells. In vitro osteogenic differentiation was most strongly and frequently observed in the RAP-derived cells, whereas rarely osteogenic differentiation was observed in the DP-derived cells. Conclusion: These findings suggest that three types of human dental tissue-derived cells from extracted wisdom teeth were multipotent mesenchymal stem cells, have the properties of multi-lineage differentiations. Especially, stem cells from root apical papilla (SCAP) have much advantage in osteogenic differentiation, whereas dental follicle cells (DFCs) have a characteristic of easy adipogenic differentiation.
Abstract
Introduction: The first aim of this study was to isolate the dental tissue-derived stem cells from the dental follicle (DF), dental pulp (DP), and root apical papilla (RAP) of the extracted wisdom teeth. Second was to evaluate their characterization with the expressions of transcription factors and cell surface markers. Finally, their ability of the in vitro multi-lineage differentiations into osteogenic and adipogenic cells were compared, respectively. Materials and Methods: Dental tissues, including dental follicle, dental pulp, and root apical papilla, were separated in the extracted wisdom teeth. These three dental tissues were cultured in Dulbecco’s modified Eagle’s medium (DMEM) with supplements, respectively. After passage 3, the homogeneous shaped dental tissue-derived cells were analyzed the expression of transcription factors (Oct-4, Nanog and Sox-2) and cell surface markers (CD44, CD90 and CD105) with reverse transcription polymerase chain reaction (RT-PCR) and fluorescence-activated cell sorting (FACS) analysis. In order to evaluate in vitro multi-lineage differentiations, the culture media were changed to the osteogenic and adipogenic induction mediums when the dental tissue-derived cells reached to passage 3. The characteristics of these three dental tissue-derived cells were compared with immunohistochemistry. Results: During primary culture, heterogenous and colony formatted dental tissue-derived cells were observed in the culture plates. After passage 2or 3, homogenous spindle-like cells were observed in all culture plates. Transcription factors and mesenchymal stem cell markers were positively observed in all three types of dental tissue-derived cells. However, the quantity of expressed transcription factors was most large in RAP-derived cells. In all three types of dental tissue-derived cells, osteogenic and adipogenic differentiations were observed after treatment of specific induction media. In vitro adipogenic differentiation was similar among these three types of cells. In vitro osteogenic differentiation was most strongly and frequently observed in the RAP-derived cells, whereas rarely osteogenic differentiation was observed in the DP-derived cells. Conclusion: These findings suggest that three types of human dental tissue-derived cells from extracted wisdom teeth were multipotent mesenchymal stem cells, have the properties of multi-lineage differentiations. Especially, stem cells from root apical papilla (SCAP) have much advantage in osteogenic differentiation, whereas dental follicle cells (DFCs) have a characteristic of easy adipogenic differentiation.
참고문헌 (36)
- Evans MJ, "Establishment in culture of pluripotential cells from mouse embryos", Nature, 292, pp.154~156 (1981)
- Wagers AJ, "Plasticity of adult stem cells", Cell, 116, pp.639~648 (2004)
- Mao JJ, "Craniofacial tissue engineering by stem cells", J Dent Res, 85, pp.966~979 (2006)
- Almeida-Porada G, "Adult stem cell plasticity and methods of detection", Rev Clin Exp Hematol, 5, pp.26~41 (2001)
- Temple S, "Stem cell plasticity-building the brain of our dreams", Nat Rev Neurosci, 2, pp.513~520 (2001)
- Yamada Y, "Bone regeneration following injection of mesenchymal stem cells and fibrin glue with a biodegradable scaffold", J Craniomaxillofac Surg, 31, pp.27~33 (2003)
- Yamada Y, "Tissue-engineered injectable bone regeneration for osseointegrated dental implants", Clin Oral Impl Res, 15, pp.589~597 (2004)
- Fuerst G, "Bone formation following sinus grafting with autogenous bone-derived cells and bovine bone mineral in minipigs: preliminary findings", Clin Oral Impl Res, 15, pp.733~740 (2004)
- Schimming R, "Tissue-engineered bone for maxillary sinus augmentation", J Oral Maxillofac Surg, 62, pp.724~729 (2004)
- Gronthos S, "Postnatal human dental pulp stem cells (DPSCs) in vitro and in vivo", Proc Natl Acad Sci USA, 97, pp.13625~13630 (2000)
- Gronthos S, "Stem cell properties of human dental pulp stem cells", J Dent Res, 81, pp.531~535 (2002)
- Miura M, "Stem cells from human exfoliated deciduous teeth", Proc Natl Acad Sci USA, 100, pp.5807~5812 (2003)
- Seo BM, "SHED repair critical-size calvarial defects in mice", Oral Dis, 14, pp.428~434 (2008)
- Huang AH, "Isolation and characterization of dental pulp stem cells from a supernumerary tooth", J Oral Pathol Med, 37, pp.571~574 (2008)
- Morsczeck C, "Isolation of precursor cells (PCs) from human dental follicle of wisdom teeth", Matrix Biol, 24, pp.155~165 (2005)
- Morsczeck C, "In vitro differentiation of human dental follicle cells with dexamethasone and insulin", Cell Biol Int, 29, pp.567~575 (2005)
- Reynolds AJ, "Cultured human and rat tooth papilla cells induce hair follicle regeneration and fiber growth", Differentiation, 72, pp.566~575 (2004)
- Seo BM, "Investigation of multipotent postnatal stem cells from human periodontal ligament", Lancet, 364, pp.149~155 (2004)
- Sonoyama W, "Mesenchymal stem cell-mediated functional tooth regeneration in swine", PLoS ONE, 1, pp.79~ (2006)
- Morsczeck C, "Comparison of human dental follicle cells (DFCs) and stem cells from human exfoliated deciduous teeth (SHED) after neural differentiation in vitro", Clin Oral Invest (2009)
- Kang EJ, "In vitro and in vivo osteogenesis of porcine skin-derived mesenchymal stem cell-like cells with a demineralized bone and fibrin scaffold", Tissue Eng Part A, 16, pp.815~827 (2010)
- Scholer HR, "New type of POU domain in germ line-specific protein Oct-4", Nature, 344, pp.435~439 (1990)
- Avilion AA, "Multipotent cell lineages in early mouse development depend on SOX2 function", Genes Dev, 17, pp.126~140 (2003)
- Chambers I, "Functional expression cloning of Nanog, a pluripotency sustaining factor in embryonic stem cells", Cell, 113, pp.643~655 (2003)
- Boyer LA, "Core transcriptional regulatory circuitry in human embryonic stem cells", Cell, 122, pp.947~956 (2005)
- Kajahn J, "Skin-derived human adult stem cells surprisingly share many features with human pancreatic stem cells", Eur J Cell Biol, 87, pp.39~46 (2008)
- Carlin R, "Expression of early transcription factors Oct-4, Sox-2 and Nanog by porcine umbilical cord (PUC) matrix cells", Reprod Biol Endocrinol, 4, pp.8~ (2006)
- Izadpanah R, "Characterization of multipotent mesenchymal stem cells from the bone marrow of rhesus macaques", Stem Cells Dev, 14, pp.440~451 (2005)
- Zhu Y, "Adipose-derived stem cell: a better stem cell than BMSC", Cell Biochem Funct, 26, pp.664~675 (2008)
- Cheng PH, "Postnatal stem", progenitor cells derived from the dental pulp of adult chimpanzee, 2008, pp.9, 20~
- Dyce PW, "Stem cells with multilineage potential derived from porcine skin", Biochem Biophys Res Commun, 316, pp.651~658 (2004)
- Zhao M, "Tracing the stemness of porcine skin-derived progenitors (pSKP) back to specific marker gene expression", Cloning Stem Cells, 11, pp.111~122 (2009)
- 최문정, "미니돼지에서 다능성 피부유래 전구세포의 추출과 이의 다배엽 세포로의 분화유도에 대한 연구", 대한구강악안면외과학회지, 34(6), pp.588~593 (2008), 대한구강악안면외과학회
- Park BW, "Osteogenic phenotypes and mineralization of cultured human periosteal-derived cells", Arch Oral Biol, 52, pp.983~989 (2007)
- Rho GJ, "Porcine mesenchymal stem cells - current technological status and future perspective", Front Biosci, 14, pp.3942~3961 (2009)
- Handa K, "Cementum matrix formation in vivo by cultured dental follicle cells", Bone, 31, pp.606~611 (2002)