Abstract:
:Somatic cell nuclear transfer (SCNT) success is partially hindered by the low epigenetic reprogramming efficiency of the donor cell. Previous studies suggest cellular heterogeneity among donor nuclei in regard to reprogramming potential, which precludes comparison among different strategies to increase cloning success. In this context, we evaluated the effect of using clonal cell populations (CPs) of bovine adult fibroblasts established by single-cell plating in SCNT. Different CPs were evaluated in regard to proliferation rate, senescence level, and chromosome stability, as well as for POU5F1 (POU class 5 homeobox 1) mRNA expression levels. In total, 9 of 24 CPs (37.5%) were successfully expanded in vitro up to the fourth passage and shown to proliferate following cryopreservation, at which time cell analyses were performed. The use of a CP with low senescence level, normal karyotype, and highest POU5F1 expression levels did not improve embryo development rates or quality following SCNT. As previously suggested, this study supports the notion that levels of POU5F1 expression in the donor nucleus do not impact the SCNT results. Notably, the single-cell seeding approach used herein to isolate CPs may be extended to the evaluation of additional predictor markers of reprogrammability success for SCNT in future experiments.
journal_name
Cell Reprogramjournal_title
Cellular reprogrammingauthors
Sá AL,Sampaio RV,da Costa Almeida NN,Sangalli JR,Brito KNL,Bressan FF,Rissino JD,do Socorro Damasceno Santos S,Meirelles FV,Ohashi OM,Dos Santos Miranda Mdoi
10.1089/cell.2016.0063subject
Has Abstractpub_date
2017-10-01 00:00:00pages
294-301issue
5eissn
2152-4971issn
2152-4998journal_volume
19pub_type
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