Rapid production and genetic stability of human mesenchymal progenitor cells derived from human somatic cell nuclear transfer-derived pluripotent stem cells

Pluripotent stem cell-derived mesenchymal progenitor cells (PSC-MPCs) are primarily derived through two main methods: 3-dimensional (3D) embryonic body-platform (EB formation) and 2D direct differentiation method. We recently established somatic cell nuclear transfer (SCNT)-PSC lines and showed their stemness. In the present study, we produced SCNT-PSC-MPCs using a novel direct differentiation method, and the characteristics, gene expression, and genetic stability of these MPCs were compared with those derived through the EB formation method. The recovery and purification of SCNT-PSC-Direct-MPCs were significantly accelerated compared to those of the SCNT-PSC-EB-MPCs, but both types of MPCs expressed typical surface markers and exhibited similar proliferation and differentiation potentials. Additionally, the analysis of gene expression patterns using microarrays showed very similar patterns. Moreover, array CGH analysis showed that both SCNT-PSC-Direct-MPCs and SCNT-PSC-EB-MPCs exhibited no significant differences in copy number variation (CNV) or single-nucleotide polymorphism (SNP) frequency. These results indicate that SCNT-PSC-Direct-MPCs exhibited high genetic stability even after rapid differentiation into MPCs, and the rate at which directly derived MPCs reached a sufficient number was higher than that of MPCs derived through the EB method. Therefore, we suggest that the direct method of differentiating MPCs from SCNT-PSCs can improve the efficacy of SCNT-PSCs applied to allogeneic transplantation. Two human SCNT (somatic cell nuclear transfer)-PSC (pluripotent stem cell) lines (CHA-SCNT-hPSC-17 and hPSC-18) were differentiated into MPCs (mesenchymal progenitor cells) using the 3D embryonic body-platform (EB formation) method (CHA-SCNT-hPSC-17-EB-MPCs and CHA-SCNT-hPSC-18-EB-MPCs) and 2D direct method (CHA-SCNT-hPSC-17-Direct-MPCs and CHA-SCNT-hPSC-18-Direct-MPCs). We performed micro-array (GeneChip® Human Gene 2.0 ST Arrays) to compare the similarity of gene expression among isogenic MPCs according to differentiation method (CHA-SCNT-hPSC-17-EB-MPC vs Direct-MPC; CHA-SCNT-hPSC-18-EB-MPC vs Direct-MPC; duplicated).