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The differential characterization of human dental progenitor cells
|關鍵字:||Stem cells from human exfoliated deciduous teeth;人類乳牙牙髓幹細胞;neurogenesis;adipogenesis;osteogenesis;chondrogenesis;神經分化;脂肪分化;硬骨分化;軟骨分化||出版社:||化學工程學系所||引用:|| Smith AG. Embryo-devrived stem cells: of mice and men. Annu Rev Cell Dev Biol 2001; 17: 435-62.  Caplan AI. Review: mesenchymal stem cells: cell-based reconstructive therapy in orthopedics. Tissue Eng 2005; 11: 1998-221.  Améen C, Strehl R, Björquist P, Lindahl A, Hyllner J, Sartipy P. Review: Human embryonic stem cells: current technologies and emerging industrial applications. Crit Rev Oncol Hematol 2008; 65: 54-80.  National Institutes of Health. Department of Health and Human Services. Stem cells: scientific progress and future research directions. 2001.http://www.nih.gov/ news/stemcell/scireport.htm  馮清榮、蔡女滿、徐偉成、韓鴻志、林欣榮。淺談幹細胞研究之展望，慈濟醫學人文月刊，2003年第2卷第1期。  台灣牙醫界，2005年24卷3期14-18。  Tuan RS, Boland G, Tuli R. Adult mesenchymal stem cells and cell-based tissue engineering. Arthritis Res Ther 2003; 5: 32-45.  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本研究比較乳牙幹細胞(stem cells from human exfoliated deciduous teeth, SHED)與其他口腔細胞(HGF)的分化特性。細胞經初代培養之後，進行細胞生長速率分析與細胞表面抗原分析；另外，在體外誘導神經、硬骨、脂肪和軟骨發生並以人類皮膚纖維母細胞(human skin fibroblasts, HSF)作為負控制組，人類骨髓間質幹細胞(human bone marrow-derived mesenchymal stem cells, HBMSC)作為正控制組。由細胞生長速率分析的結果發現，HGF和SHED兩種細胞的倍增時間分別為65.2±3.1小時和51.2±4.4小時，HGF和SHED在CD29、CD44、CD73、CD90和CD105表面抗原呈陽性，與人類骨髓間質幹細胞具有相同的表面抗原表現。HGF和SHED經體外誘導神經分化4.5天後形態明顯改變；21天後神經特異性的基因和蛋白質都明顯增加。經低細胞密度誘導硬骨分化21天後，RUNX-2和鹼性磷酸酶的表現量增加，而在硬骨分化晚期所表現的成熟基因osteocalcin，亦表現出成熟硬骨細胞的礦化現象。以高細胞密度誘導脂肪分化21天後，脂肪特異性基因PPARg2和LPL的表現量增加，同時有成熟脂肪細胞油滴的累積。以三度空間細胞微珠誘導軟骨發生21天後，能使軟骨分化基因的表現量增加且有大量的軟骨基質分泌。以上實驗證實HGF和SHED具有多向分化的能力，其分化特性不輸於HBMSC，而HGF的分化能力也與SHED相似，甚至在軟骨分化上，HGF更有潛力。本研究中，發現口腔前驅細胞能分化成中胚層的硬骨細胞、脂肪細胞和軟骨細胞，亦具有跨胚層分化成外胚層的神經細胞的能力，表現與骨髓間質幹細胞相似，相信在未來將可做為另一個成體幹細胞的來源。
This study investigates whether human dental tissue contains a certain population of cells that have intrinsic differentiation potential. Cells derived from dental tissue were characterized by FACS-analysis and cultured for 21 days separately in the osteogenic, adipogenic, chondrogenic and neurogenic media. Phenotypical evaluation was carried out by immunohistochemical analysis and RT-PCR analysis. The results were compared with those from adult bone marrow-derived stromal cells. Dental tissue-derived cells were negative for CD34, CD31, CD45, and were positive for CD29, CD44, CD73, CD90, and CD105. In the adipogenic medium, the intracellular lipid vacuoles were visible and dental tissue-derived cells showed upregulation of adipogenic marker genes LPL and PPARg2. In the chondrogenic medium, gingiva-derived cells were positively stained for collagen type II and showed upregulation of both collagen type II and sox9 genes. In the osteogenic medium, von Kossa staining showed calcium deposition and cells showed upregulation of OCN and RUNX2 genes. In the neurogenic medium, cells showed upregulation of GFAP and beta-III tubulin both at mRNA and protein levels. This study suggests that gingiva-derived cells may have an intrinsic differentiation potential. Gingiva-derived cells generated both neural and mesodermal progeny. From this research we propose that gingiva-derived cells represent a population of novel multipotent adult stem cells and may provide an accessible and autologous source of stem cells for transplantation.
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