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Upper, passage 3; lower, passage 9. Having shown the distinct senescence-associated kinetics in HGPS-MSCs and WS-MSCs, we continued to evaluate other aging-related phenotypes.
By using lower passage (numbers 3 5) EG cells as donor cells, the SCNT efficiency was significantly lower compared with use of fetal fibroblast donor cells.
Lower passage SMC adopted a contractile phenotype marked by a relatively slower proliferation rate, higher expression of proteins of the contractile apparatus and smoothelin, elongated morphology, and reduced deposition of collagen types I and III.
Most interestingly, these extensively passaged P7 ACs (expansion factor of 85,000), which are typically highly dedifferentiated, were able to form constructs with properties similar to or higher than those formed by lower passage number cells.
In addition, the frequency of sex chromosome loss was higher in higher passage than in lower passage cells as demonstrated for other chromosomal aberrations in HESCs [28], [29].
Indeed, we found that two ARO cultures that had been passaged 50 or more times contained a higher proportion (∼50.1%) of CD133+ cells than did ARO cultures of lower passage numbers (∼21.9%)(Fig. 3B).
Similar(49)
Results showed that genes either associated with DNA demethylation including DNMTs and TET1 or genes related to histone acetylation including HDACs were highly expressed in EG cells at higher passages when compared to EG cells at lower passages.
Here, among the debris of broken pottery and shattered stone vessels from the burial chamber and lower passages were the remnants of two vases made for Queen Ahmose Nefertari which formed part of the original funerary equipment of Thutmose I; one of the vases contained a secondary inscription which states that Thutmose II "[made it] as his monument to his father".
Subconfluent cultures (70 90% confluence) of lower passages (below passage 9) were utilized for the experiments described.
Interestingly, expression of adipogenic markers was significantly stronger in higher passages of T-MPCs (P4 and P5) than in lower passages (P2 and P3) (data not shown).
The cellular immortalization resulted in a greater expansion of osteoarthritic chondrocytes which under normal physiological conditions would become senescent at lower passages.
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