Comparative Analysis of Nuclear Transfer Embryo-Derived Mouse Embryonic Stem Cells. Part I: Cellular Characterization

Abstract

Embryonic stem cells derived from nuclear transfer embryos (ntESCs) are particularly valuable for regenerative medicine, as they are a patient-specific and histocompatible cell source for the treatment of varying diseases. However, currently, little is known about their cellular and molecular profile. In the present study, in a mouse model different donor cell-derived ntESCs from various genetic backgrounds were compared with reference ESCs and analyzed comprehensively at the cellular level. A number of pluripotency marker genes were compared by flow cytometry and immunocytochemistry analysis. Significant differences at the protein level were observed for POU5F1, SOX2, FGF4, NANOG, and SSEA-1. However, such differences had no effect on in vitro cell differentiation and cell fate: derivatives of the three germ layers were detected in all ntESC lines. The neural and cardiac in vitro differentiation revealed minor differences between the cell lines, both at the mRNA and protein level. Karyotype analyses and cell growth studies did not reveal any significant variations. Despite some differences observed, the present study revealed that ntESC lines had similar differentiation competences compared to other ESCs. The results indicate that the observed differences may be related to the genotype rather than to the nuclear transfer technology.

Slow Turning Lateral Vessel Bioreactor Improves Embryoid Body Formation and Cardiogenic Differentiation of Mouse Embryonic Stem Cells

“…This is in agreement with our observations of a low efficiency of HD-EBs to differentiate into cardiac cells, when plated later (D4 and D5). In standard cardiac differentiation protocols using HD culture, the initial cell seeding density is usually 400-1000 ESCs/mL, and the EBs are usually plated on gelatin-coated dishes on D5 (Chen et al, 2011;Kobolak et al, 2012;Mummery et al, 2002;Passier and Mummery, 2005), resulting a general low efficiency of cardiac differentiation. This may be due to the mistiming in plating of the EBs onto gelatin.…”

Allele-specific RNA-seq expression profiling of imprinted genes in mouse isogenic pluripotent states

“…S18a). As female ESCs are prone to the loss of one X chromosome, we analyzed the karyotype of the ESC lines as a control (Additional file 1: Table S1; [61]). This showed that the ESC-NT2 line is largely X0, explaining the allelic bias and the reduction in X-linked gene expression for ESC-NT2 ( Fig.…”

Lymphoid lineage differentiation potential of mouse nuclear transfer embryonic stem cells

“…Rejection by host body immune system due to incompatibility of MHC types is considered as the most important hurdle in cell therapy [5]. Several attempts have been made to tackle such problem among which using the cells with the same MHC as reception such as nuclear transfer embryonic stem cells (nt-ESCs) and induced pluripotent stem cells (iPSCs) are the most efficient strategies [6]. Nuclear transfer process enables reprogramming of somatic cell nuclei and its transformation to embryonic cells [7].…”

“…Recent studies reported some differences in characteristics of ESCs and iPSCs. On the other hand, differentiated cells derivediPSCs show more sensitivity to apoptosis and less proliferation rate in comparison with nt-ESCs and ESCs [6,10]. Possessing the same MHC to the donor somatic cell, as a distinctive characteristic of the nt-ESCs, has made these cells a potential source for stem cell therapy and regenerative medicine [9,11].…”