Species‐specific Challenges in Dog Cloning

Kim, G A; Oh, Hyun Joo; Park, J E; Kim, M J; Park, E J; Jo, Young-Kwang; Jang, Goo; Kim, M K; Kim, H J; Lee, B C

doi:10.1111/rda.12035

Cited by 5 publications

(4 citation statements)

References 41 publications

(70 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In the present study, OCT4 expression in ASC and FF was significantly higher than in AF, and SOX2 expression was the highest in ASC. In agreement with our results, OCT4 is expressed in bone marrow MSC, equine cord blood cells and canine ASC (Reed & Johnson 2008;Violini et al 2009;Kim et al 2012). It is believed that high expression levels of genes like OCT4 and SOX2 may be necessary for stemness regulation of porcine ASC.…”

Section: Discussionsupporting

confidence: 91%

See 1 more Smart Citation

Analysis of cell growth and gene expression of porcine adipose tissue‐derived mesenchymal stem cells as nuclear donor cell

Park

et al. 2014

Dev Growth Differ

View full text Add to dashboard Cite

In several laboratory animals and humans, adipose tissue-derived mesenchymal stem cells (ASC) are of considerable interest because they are easy to harvest and can generate a huge proliferation of cells from a small quantity of fat. In this study, we investigated: (i) the expression patterns of reprogramming-related genes in porcine ASC; and (ii) whether ASC can be a suitable donor cell type for generating cloned pigs. For these experiments, ASC, adult skin fibroblasts (AF) and fetal fibroblasts (FF) were derived from a 4-year-old female miniature pig. The ASC expressed cell-surface markers characteristic of stem cells, and underwent in vitro differentiation when exposed to specific differentiation-inducing conditions. Expression of DNA methyltransferase (DNMT)1 in ASC was similar to that in AF, but the highest expression of the DNMT3B gene was observed in ASC. The expression of OCT4 was significantly higher in FF and ASC than in AF (P < 0.05), and SOX2 showed significantly higher expression in ASC than in the other two cell types (P < 0.05). After somatic cell nuclear transfer (SCNT), the development rate of cloned embryos derived from ASC was comparable to the development of those derived using FF. Total cell numbers of blastocysts derived using ASC and FF were significantly higher than in embryos made with AF. The results demonstrated that ASC used for SCNT have a potential comparable to those of AF and FF in terms of embryo in vitro development and blastocyst formation.

show abstract

Section: Discussionsupporting

confidence: 91%

“…; Kim et al . ). It is believed that high expression levels of genes like OCT4 and SOX2 may be necessary for stemness regulation of porcine ASC.…”

Section: Discussionmentioning

confidence: 97%

Analysis of cell growth and gene expression of porcine adipose tissue‐derived mesenchymal stem cells as nuclear donor cell

Park

et al. 2014

Dev Growth Differ

View full text Add to dashboard Cite

show abstract

“…Canine in vivo matured oocytes were recovered by aseptic surgical procedures 70–76 h after the day of ovulation which was considered as the day when the serum progesterone concentration reached 4.0–9.9 ng/mL as previously described (Kim et al . ). Oocytes surrounded by cumulus cell layers were denuded by repeated pipetting in HEPES‐buffered tissue culture medium (TCM)‐199 supplemented with 0.1% (w/v) hyaluronidase.…”

Section: Methodsmentioning

confidence: 97%

Propagation of elite rescue dogs by somatic cell nuclear transfer

Choi

Kim

et al. 2015

Animal Science Journal

View full text Add to dashboard Cite

The objective of the present study was to compare the efficiency of two oocyte activation culture media to produce cloned dogs from an elite rescue dog and to analyze their behavioral tendencies. In somatic cell nuclear transfer procedure, fused couplets were activated by calcium ionophore treatment for 4 min, cultured in two media: modified synthetic oviduct fluid (mSOF) with 1.9 mmol/L 6-dimethylaminopyridine (DMAP) (SOF-DMAP) or porcine zygote medium (PZM-5) with 1.9 mmol/L DMAP (PZM-DMAP) for 4 h, and then were transferred into recipients. After embryo transfer, pregnancy was detected in one out of three surrogate mothers that received cloned embryos from the PZM-DMAP group (33.3%), and one pregnancy (25%) was detected in four surrogate mothers receiving cloned embryos from the SOF-DMAP group. Each pregnant dog gave birth to one healthy cloned puppy by cesarean section. We conducted the puppy aptitude test with two cloned puppies; the two cloned puppies were classified as the same type, accepting humans and leaders easily. The present study indicated that the type of medium used in 6-DMAP culture did not increase in cloning efficiency and dogs cloned using donor cells derived from one elite dog have similar behavioral tendencies.

show abstract

“…The PA zygotes were used in 2 experiments: (1) observing DNA synthesis by BrdU incorporation at 2, 4, and 8 h post-DMAP treatment (hpd), and (2) confirming fetus formation after ET. After DMAP treatment, the PA zygotes were immediately transferred surgically to the oviducts of recipients in which estrus was naturally synchronized as previously described [ 35 , 36 ]. Synchronous recipients (where ovulation on the same day as the oocyte donor) were identified by monitoring serum progesterone concentration.…”

Section: Methodsmentioning

confidence: 99%

Optimal Treatment of 6-Dimethylaminopurine Enhances the In Vivo Development of Canine Embryos by Rapid Initiation of DNA Synthesis

Lee

Kim

2021

IJMS

View full text Add to dashboard Cite

Artificial activation of oocytes is an important step for successful parthenogenesis and somatic cell nuclear transfer (SCNT). Here, we investigated the initiation of DNA synthesis and in vivo development of canine PA embryos and cloned embryos produced by treatment with 1.9 mM 6-dimethylaminopurine (6-DMAP) for different lengths of time. For experiments, oocytes for parthenogenesis and SCNT oocytes were cultured for 4 min in 10 μM calcium ionophore, and then divided into 2 groups: (1) culture for 2 h in 6-DMAP (DMAP-2h group); (2) culture for 4 h in DMAP (DMAP-4h group). DNA synthesis was clearly detected in all parthenogenetic (PA) embryos and cloned embryos incorporated BrdU 4 h after activation in DMAP-2h and DMAP-4h groups. In vivo development of canine parthenogenetic fetuses was observed after embryo transfer and the implantation rates of PA embryos in DMAP-2h were 34%, which was significantly higher than those in DMAP-4h (6.5%, p < 0.05). However, in SCNT, there was no significant difference in pregnancy rate (DMAP-2h: 41.6% vs. DMAP-4h: 33.3%) and implantation rates (DMAP-2h: 4.94% vs. DMAP-4h: 3.19%) between DMAP-2h and DMAP-4h. In conclusion, the use of DMAP-2h for canine oocyte activation may be ideal for the in vivo development of PA zygotes, but it was not more effective in in vivo development of canine reconstructed SCNT oocytes. The present study demonstrated that DMAP-2h treatment on activation of canine parthenogenesis and SCNT could effectively induce the onset of DNA synthesis during the first cell cycle.

show abstract

Species‐specific Challenges in Dog Cloning

Cited by 5 publications

References 41 publications

Analysis of cell growth and gene expression of porcine adipose tissue‐derived mesenchymal stem cells as nuclear donor cell

Analysis of cell growth and gene expression of porcine adipose tissue‐derived mesenchymal stem cells as nuclear donor cell

Propagation of elite rescue dogs by somatic cell nuclear transfer

Optimal Treatment of 6-Dimethylaminopurine Enhances the In Vivo Development of Canine Embryos by Rapid Initiation of DNA Synthesis

Contact Info

Product

Resources

About