direct differentiation method
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2021 ◽  
Vol 22 (17) ◽  
pp. 9238
Author(s):  
Soo Kyung Jung ◽  
Jeoung Eun Lee ◽  
Chang Woo Lee ◽  
Sung Han Shim ◽  
Dong Ryul Lee

Pluripotent stem cell-derived mesenchymal progenitor cells (PSC-MPCs) are primarily derived through two main methods: three-dimensional (3D) embryoid body-platform (EB formation) and the 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 EB formation. 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.


Author(s):  
Victor Makhniy ◽  
Oksana Kinzerska ◽  
Illia Senko

A comparative analysis of various methods for the decomposition of broad bands into individual components has been carried out. It is shown that the most universal are the methods of modulation spectroscopy and direct differentiation of conventional spectral characteristics, which, unlike the widely used Alentsev-Fock method, are applicable to spectra of any type (luminescence, transmission, absorption, etc.). The features and capabilities of the direct differentiation method are used to identify the structure in the emission spectra and transmission of ZnSe <Al> single-crystal substrates.


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