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        The differentiation of rat-induced pluripotent stem cells into alveolar type II epithelial cells with a three-step induction protocol

        Bei Wu,Chen Wang,Feilong Hei,Cun Long,Mengmeng Chen,Shengnan Yang,Jie Yu,Zhihai Ju 한국통합생물학회 2015 Animal cells and systems Vol.19 No.1

        Induced pluripotent stem (iPS) cells derive from autologous somatic cells, the application prospect of iPS cells forregenerative medicine and tissue engineering is better than embryonic stem cells (ESCs) to some extent. Alveolar type II(AT II) epithelial cells play key role in the injured lung tissue regeneration and function recovery. The differentiation of iPScells into AT II cells could provide available source for injured lung treatment. In this study, rat iPS (riPS) cells wereresuscitated and proliferated for 14 days before differentiation. A modified three-step induction protocol similar to thereported ESCs inducing procedure was used in this study for the differentiation groups. Routine cell culture was done to theriPS cell control group (riPS-con). At stage 3, cells of day 7 (Diff. 7) and day 14 (Diff. 14) were collected for the real-timepolymerase chain reaction tests for gene expressions of Oct4, Nanog, SPA, SPB, SPC, SPD, and CC10. Immunofluorescencestaining of SPC and SSEA-1 was conducted. At the end of the differentiation, cell morphology becameoutstretched and epithelium-like. Cells of the Diff. 14 group positively expressed SPC and negatively expressed SSEA-1,which is contrary to the riPS-con group. In the Diff. 7 and the Diff. 14 groups, the expression of Oct4, Nanog, and SPBdecreased (P < 0.05), whereas the expression of SPA, SPC, SPD (P < 0.05), and CC10 (P > 0.05) increased. This studyindicated that riPS cells can successfully differentiate into AT II epithelial cells with the three-step induction protocol andmay be further applied to implanting in decellularized rat lung scaffolds and building a bio-artificial lung.

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