Increasing maternal age of blastocyst affects on efficient derivation and behavior of mouse embryonic stem cells

Abstract

Mouse embryonic stem cells (mESCs) have the capability to undergo unlimited cell division and differentiate into derivatives of all three embryonic germ layers. These fundamental features enable mESCs to potentially be appropriate, efficient models for biological and medical research. Therefore, it is essential to produce high‐performance mESCs. In the current study, we have produced mESCs from blastocysts that developed from fertilized oocytes of 2 (2‐C57)‐, 4 (4‐C57)‐, and 6 (6‐C57)‐month‐old C57BL/6 mice. A comparison of isolated stem cells was done from the viewpoint of the efficiency of mESC derivation, self‐renewal, and their differentiation capacity. All generated mESCs showed a similar expression of the molecular markers protein of pluripotency and AP activity. In the 3i medium, there was a significant decrease in undifferentiated marker genes expression in the 2‐C57 cells compared with the other two groups ( P\u2009<\u20090.05) but developmental genes significantly increased in the 4‐C57 and 6‐C57 cells compared with the 2‐C57 cells ( P\u2009<\u20090.05). The differentiation capacity into three germ layers through the embryoid body formation and percentage of cell lines with normal numbers of chromosomes reduced with increased maternal age. The highest DT and highest percentage of cells in the S phase belonged to 2‐C57 cells. These data demonstrated that blastocysts which developed from fertilized oocytes of 2‐, 4‐, and 6‐month‐old C57BL/6 mice can generate pluripotent stem cells, and suggested that both the efficiency of mESC isolation and the behavior of these isolated mESCs including pluripotency, self‐renewal, cell cycle, and DT changed with increasing maternal age.