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The persistence of A-type nuclear lamin in somatic cell nuclear transfer (SCNT) embryos has been proposed as a marker for incomplete nuclear reprogramming. Using monoclonal antibodies to A/C- (A/C-346 and A/C-131C3) and B-type lamin, we compared distribution during early development of bovine IVF, parthenogenetic and SCNT embryos. A/C-346 staining was observed in the pronuclei of IVF embryos and in nuclei at the two-cell stage, but was not detected in subsequent cleavage stages up to and including hatched blastocysts. In contrast, A/C-131C3 and anti-lamin B2 stained all preimplantation stage embryos. Parthenogenetic and SCNT embryos had similar staining patterns to IVF embryos for all three antibodies, demonstrating correct nuclear architecture reprogramming. Inhibiting protein synthesis with cycloheximide (CHX) in parthenogenetic and SCNT embryos did not affect lamin A/C localisation, suggesting that lamin A/C is maternal in origin. However, activation with CHX delayed lamin A/C incorporation compared with 6-dimethylaminopurine activation. In SCNT embryos, staining for both A/C- and B-type lamin was delayed compared with parthenotes, although lamin B2 incorporation preceded lamin A/C in both. In conclusion, the lamin A/C distribution in SCNT bovine embryos paralleled that of IVF and parthenogenetic controls and therefore is not a marker of incomplete reprogramming.
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http://dx.doi.org/10.1071/RD09264 | DOI Listing |
Reprod Domest Anim
September 2025
National Engineering Research Center for Breeding Swine Industry, South China Agricultural University, Guangzhou, China.
Canine somatic cell nuclear transfer (SCNT) is a powerful technology that can be used to clone beloved companion dogs, produce valuable working dogs, rescue endangered canine breeds, and create genetically engineered dogs. Nevertheless, the application of this technology is hindered by the low developmental efficiency of canine SCNT embryos. It has been shown that in pig and horse cloning using mesenchymal stem cells (MSCs), compared with fibroblasts, as donor cells can enhance the developmental potential of SCNT embryos.
View Article and Find Full Text PDFTheriogenology
December 2025
Guangxi Key Laboratory of Animal Breeding & Disease Control and Prevention, Guangxi University, 75 Xiuling Road, Nanning, Guangxi, 530005, China. Electronic address:
The molecular mechanisms underlying differences in somatic cell cloning efficiency among buffalo fetal fibroblasts (BFFs) from different individuals remain poorly understood. Our study conducted a comparative analysis of mitochondrial function and chromatin remodeling capacity of BFFs from different individuals, and tracked the mitochondria in the embryos derived from SCNT and IVF. These findings revealed that BFFs with high cloning efficiency displayed well-preserved mitochondrial morphology and ultrastructure, increased mitochondrial DNA copy number and ATP levels, elevated antioxidant capacity, enhanced mitochondrial membrane potential, and significantly upregulated expression levels of mitochondria-related genes.
View Article and Find Full Text PDFFront Cell Dev Biol
July 2025
Division of Applied Life Science (BK21 Four), Gyeongsang National University, Jinju, Gyeongnam, Republic of Korea.
Somatic cell nuclear transfer (SCNT) is important in assisted reproductive technologies. However, its reprogramming efficiency remains low. A considerable drawback of SCNT-cloned embryos is the reduction in telomerase activity, which is crucial for DNA stability and genetic and epigenetic reprogramming.
View Article and Find Full Text PDFSci Rep
July 2025
Embryo Biotechnology Lab, Animal Biotechnology Division (ABTD), ICAR- National Dairy Research Institute, Karnal, 132001, Haryana, India.
Somatic cell nuclear transfer (SCNT) allows the multiplication of elite livestock and conservation of endangered species. However, restrictions on cow slaughter limit the access to oocytes for SCNT applications in Indian cattle breeds. To overcome this limitation, we utilized transvaginal ovum pick-up (OPU) method to collect oocytes, which were then used for the production of cloned embryos via the handmade cloning (HMC) technique.
View Article and Find Full Text PDFReprod Biol
September 2025
Kheiron S.A., Pilar, Buenos Aires B1629, Argentina. Electronic address:
This study aimed to obtain myostatin (MSTN)-knockout calves, while avoiding the risk of exogenous DNA integration during gene editing. To achieve this, we combined CRISPR/Cas9 ribonucleoprotein gene editing with somatic cell nuclear transfer (SCNT) technology. In the first experiment (E1), we compared the gene editing efficiency of four gRNAs targeting different coding regions of the MSTN gene using plasmid-based CRISPR/Cas9 in bovine fetal fibroblasts (BFF-E1 cells).
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