Differentiation of mouse embryonic stem cells into dental epithelial-like cells induced by ameloblasts serum-free conditioned medium.
Embryonic stem cells (ESCs) possess an intrinsic self-renewal ability and can differentiate into numerous types of functional tissue cells; however, whether ESCs can differentiate toward the odontogenic lineage is still unknown. In this study, we developed an efficient culture strategy to induce the differentiation of murine ESCs (mESCs) into dental epithelial cells. By culturing mESCs in ameloblasts serum-free conditioned medium (ASF-CM), we could induce their differentiation toward dental epithelial cell lineages; however, similar experiments with the tooth germ cell-conditioned medium (TGC-CM) did not yield effective results. After culturing the cells for 14days in the differentiation-inducing media, the expression of ameloblast-specific proteins such as cytokeratin (CK)14, ameloblastin (AMBN), and amelogenin (AMGN) was markedly higher in mESCs obtained with embryoid body (EB) formation than in mESCs obtained without EB formation. We observed that immunocompromised mice implanted with induced murine EBs (mEBs) showed tissue regenerative capacity and produced odontogenic epithelial-like structures, whereas those implanted with mSCE monolayer cells mainly formed connective tissues. Thus, for the first time, we report that ASF-CM provides a suitable microenvironment for inducing mESC differentiation along the odontogenic epithelial cell lineage. This result has important implications for tooth tissue engineering.
Ning
Fang
F
Department of Orthodontics, School of Stomatology, Fourth Military Medical University, Xi'an, 710032 Shaanxi, People's Republic of China.
Guo
Yunshan
Y
Tang
Juan
J
Zhou
Jing
J
Zhang
Hongmei
H
Lu
Wei
W
Gao
Yuan
Y
Wang
Lei
L
Pei
Duanqing
D
Duan
Yinzhong
Y
Jin
Yan
Y
D000565:Ameloblasts / Q000378:metabolism*; D053523:Amelogenin / Q000096:biosynthesis; D000818:Animals; D002460:Cell Line; D019070:Cell Lineage; D017077:Culture Media, Conditioned / Q000378:metabolism / Q000494:pharmacology; D016895:Culture Media, Serum-Free / Q000378:metabolism / Q000494:pharmacology; D003746:Dental Enamel Proteins / Q000096:biosynthesis; D053595:Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects*; D004847:Epithelial Cells / Q000166:cytology; D053547:Keratin-14 / Q000096:biosynthesis; D051379:Mice; D009805:Odontogenesis; D023822:Tissue Engineering / Q000379:methods*; D014070:Tooth / Q000166:cytology / Q000254:growth & development*
C490890:Ambn protein, mouse; D053523:Amelogenin; D017077:Culture Media, Conditioned; D016895:Culture Media, Serum-Free; D003746:Dental Enamel Proteins; D053547:Keratin-14