Expression of Multiple Stem Cell Markers in Dental Pulp Cells Cultured in Serum-free Media

被引:33
作者
Hirata, Thais Miyuki [1 ]
Ishkitiev, Nikolay [1 ]
Yaegaki, Ken [1 ,2 ]
Calenic, Bogdan [1 ]
Ishikawa, Hiroshi [3 ]
Nakahara, Taka [3 ]
Mitev, Vanyo [4 ]
Tanaka, Tomoko [1 ]
Haapasalo, Markus [2 ]
机构
[1] Nippon Dent Univ Tokyo, Dept Oral Hlth, Sch Life Dent Tokyo, Chiyoda Ku, Tokyo, Japan
[2] Univ British Columbia, Dept Oral Biol & Med Sci, Vancouver, BC V5Z 1M9, Canada
[3] Nippon Dent Univ Tokyo, Sch Life Dent Tokyo, Sect Dev & Regenerat Dent, Tokyo, Japan
[4] Med Univ Sofia, Dept Med Chem & Biochem, Sofia, Bulgaria
关键词
Culture media; dental pulp; ectoderm; endoderm; mesoderm; serum-free; stem cell; stem cell transplantation; FIBROBLAST-GROWTH-FACTOR; MARROW STROMAL CELLS; FEEDER-FREE; IN-VITRO; DIFFERENTIATION; PROLIFERATION; LINES; VIVO;
D O I
10.1016/j.joen.2010.03.002
中图分类号
R78 [口腔科学];
学科分类号
1003 ;
摘要
Introduction: Stem cell lines are usually grown in medium containing animal products. Fetal bovine serum (FBS) is an important additive for cell growth; however, the allergenic potential and the possibility of contamination when we use a medium containing serum would be a barrier to transplantation and consequently to the introduction of cell therapy methods into clinical applications. Methods: Dental mesenchymal cells were isolated and expanded in vitro and maintained in 4 different serum-free media (SFMs): SFM#1 (ITS-X, embryotrophic factor [ETF]); SFM#2 (ITS-X); SFM#3 (ETF); and SFM#4 (ETF, sodium pyruvate, ascorbic acid, fibroblast growth factor [FGF-a], acidic). Viability, proliferative, and immunocytochemical tests for the cells were performed by using 4 stem cell markers (CD44H, CK19, nestin, and P63) for ectoderm, mesoderm, and endoderm. Results: Viability tests showed a significant difference between the control and SFMs in both deciduous tooth pulp cells (DTPCs) and wisdom tooth pulp cells (WTPCs). However, all SFMs demonstrated 84%-90% viability, whereas the control showed 90%-93%. In both DTPCs and WTPCs, SFM#1 had the highest proliferation rate among the 4 SFMs. Immunocytochemistry stained positive stem cell markers most intensely in cells cultured with SFM#1. Furthermore, all stem cell markers for ectoderm, mesoderm, and endoderm were expressed in the cells cultured with SFM#1. Conclusions: SFM#1 showed an acceptable survival rate, the highest proliferation rate, and the strongest expression of all the stem cell markers. SFM#1 proved to be a suitable medium for the culture of human dental pulp stem cells and to preserve pluripotency in differentiation. (J Endod 2010;36:1139-1144)
引用
收藏
页码:1139 / 1144
页数:6
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