Mycoplasma detection and elimination are necessary for the application of stem cell from human dental apical papilla to tissue engineering and regenerative medicine.
BACKGROUND
Recently, postnatal stem cells from dental papilla with neural crest origin have been considered as one of potent stem cell sources in regenerative medicine regarding their multi-differentiation capacity and relatively easy access. However, almost human oral tissues have been reported to be infected by mycoplasma which gives rise to oral cavity in teeth, and mycoplasma contamination of ex-vivo cultured stem cells from such dental tissues and its effect on stem cell culture has received little attention.


RESULTS
In this study, mycoplama contamination was evaluated with stem cells from apical papilla which were isolated from human third molar and premolars from various aged patients undergoing orthodontic therapy. The ex-vivo expanded stem cells from apical papilla were found to express stem cell markers such as Stro-1, CD44, nestin and CD133, but mycoplama contamination was detected in almost all cell cultures of the tested 20 samples, which was confirmed by mycoplasma-specific gene expression and fluorescence staining. Such contaminated mycoplasma could be successfully eliminated using elimination kit, and proliferation test showed decreased proliferation activity in mycoplasma-contaminated cells. After elimination of contaminated mycoplasma, stem cells from apical papilla showed osteogenic and neural lineage differentiation under certain culture conditions.


CONCLUSIONS
Our study proposes that the evaluation of mycoplasma contamination and elimination process might be required in the use of stem cells from apical papilla for their potent applications to tissue engineering and regenerative medicine.
Department of Maxillofacial Biomedical Engineering and Institute of Oral Biology, School of Dentistry, Kyung Hee University, 1 Hoegi-dong, Dongdaemun-gu, Seoul, 130-701 Republic of Korea.
Byung-Chul
BC
Kim
Department of Maxillofacial Biomedical Engineering and Institute of Oral Biology, School of Dentistry, Kyung Hee University, 1 Hoegi-dong, Dongdaemun-gu, Seoul, 130-701 Republic of Korea.
So Yeon
SY
Kim
Department of Oral Maxillofacial Surgery, School of Dentistry, Kyung Hee University, Seoul, 130-701 Republic of Korea.
Yong-Dae
YD
Kwon
Department of Pediatric Dentistry, School of Dentistry, Kyung Hee University, Seoul, 130-701 Republic of Korea.
Sung Chul
SC
Choe
Department of Cogno-Mechatronics Engineering, Pusan National University, Busan, 609-735 Korea.
Dong-Wook
DW
Han
Department of Maxillofacial Biomedical Engineering and Institute of Oral Biology, School of Dentistry, Kyung Hee University, 1 Hoegi-dong, Dongdaemun-gu, Seoul, 130-701 Republic of Korea.
Yu-Shik
YS
Hwang
Dental papilla; Differentiation; Mycoplasma; Neural; Osteogenic; Proliferation; Stem cell