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. 2014 Jan 22;9(1):e86011. doi: 10.1371/journal.pone.0086011

Figure 1. Protocol for tooth organ engineering.

Figure 1

The mandibular first molars were dissected from ICR mouse embryos at embryonic day (ED) 14 (cap stage) (A). Then, the dental epithelium (B) and ecto-mesenchyme (C) were separated by using a mixture of 0.25% trypsin and 1.2 U/mL dispase in DMEM-F12 (preheated to 37°C) at room temperature during 15 min. Each tissue was dissociated into single cells, which were then re-associated (D) and grown on semi-solid cultured medium (E). After 7 days in vitro, each re-association was co-cultured overnight with trigeminal ganglia from ICR newborn mice (F). The eighth day (G), bioengineered tooth unit and trigeminal ganglia were co-implanted between skin and muscles behind the ears in adult ICR mice (mice a), CsA-treated ICR mice (mice b) and Nude mice (mice c) for 1 week or 2 weeks (H). BT, bioengineered tooth; TG, trigeminal ganglia.