Osteo-/odontogenic differentiation of induced mesenchymal stem cells generated through epithelial-mesenchyme transition of cultured human keratinocytes.
Osteo-/odontogenic differentiation of induced mesenchymal stem cells generated through epithelial-mesenchyme transition of cultured human keratinocytes.
复制标题
通过培养的人角质形成细胞的上皮-间质转化产生的诱导间充质干细胞的骨/牙源性分化。
DOI：
10.1016/j.joen.2014.07.014
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发表时间：
2014
影响因子：
4.2
通讯作者：
Kang,MoK
中科院分区：
文献类型：
--
作者：
Yi,Jin-Kyu;Mehrazarin,Shebli;Oh,Ju-Eun;Bhalla,Anu;Oo,Jenessa;Chen,Wei;Lee,Min;Kim,ReubenH;Shin,Ki-Hyuk;Park,No-Hee;Kang,MoK
IntroductionRevascularization of necrotic pulp has been successful in the resolution of periradicular inflammation; yet, several case studies suggest the need for cell-based therapies using mesenchymal stem cells (MSCs) as an alternative for de novo pulp regeneration. Because the availability of MSCs may be limited, especially in an aged population, the current study reports an alternative approach in generating MSCs from epidermal keratinocytes through a process called epithelial-mesenchymal transition (EMT).MethodsWe induced EMT in primary normal human epidermal keratinocytes (NHEKs) by transient transfection of small interfering RNA targeting thep63gene. The resulting cells were assayed for their mesenchymal marker expression, proliferation capacities as a monolayer and in a 3-dimensional collagen scaffold, and differentiation capacities.ResultsTransient transfection ofp63small-interfering RNA successfully abolished the expression of endogenousp63in NHEKs and induced the expression of mesenchymal markers (eg, vimentin and fibronectin), whereas epithelial markers (eg, E-cadherin and involucrin) were lost. The NHEKs exhibiting the EMT phenotype acquired extended replicative potential and an increased telomere length compared with the control cells. Similar to the established MSCs, the NHEKs withp63knockdown showed attachment onto the 3-dimensional collagen scaffold and underwent progressive proliferation and differentiation. Upon differentiation, these EMT cells expressed alkaline phosphatase activity, osteocalcin, and osteonectin and readily formed mineralized nodules detected by alizarin S red staining, showing osteo-/odontogenic differentiation.ConclusionsThe induction of EMT in primary NHEKs by means of transientp63knockdown allows the generation of induced MSCs from autologous sources. These cells may be used for tissues engineering purposes, including that of dental pulp.
