Update on the First Cloned Dog and Outlook for Canine Cloning

2015 ◽  
Vol 17 (5) ◽  
pp. 325-326
Author(s):  
Goo Jang ◽  
ByeongChun Lee
Keyword(s):  
Cloned Dog

Desperately Seeking Snuppy’s Mom(s) : Re-writing the Story of the First-Cloned Dog

2016 ◽  
Vol 16 (1) ◽  
pp. 3
Author(s):  
Ji-Eun Lee
Keyword(s):  
Cloned Dog

The promise of dog cloning

2018 ◽  
Vol 30 (1) ◽  
pp. 1 ◽  
Author(s):  
Hyun Ju Oh ◽  
Kihae Ra ◽  
Min Jung Kim ◽  
Geon A Kim ◽  
Erif Maha Nugraha Setyawan ◽  
...  
Keyword(s):  
Somatic Cell ◽  
Somatic Cell Nuclear Transfer ◽  
Nuclear Transfer ◽  
Current Method ◽  
The World ◽  
Cloned Dog

Dog cloning as a concept is no longer infeasible. Starting with Snuppy, the first cloned dog in the world, somatic cell nuclear transfer (SCNT) has been continuously developed and used for diverse purposes. In this article we summarise the current method for SCNT, the normality of cloned dogs and the application of dog cloning not only for personal reasons, but also for public purposes.

37 RESTORING REPROGRAMMING ABNORMALITIES IN A CLONED DOG HAVING ECTOPIC LIVER AND GALL BLADDER BY RECLONING

2014 ◽  
Vol 26 (1) ◽  
pp. 133
Author(s):  
M. J. Kim ◽  
H. J. Oh ◽  
G. A. Kim ◽  
Y. K. Jo ◽  
J. Choi ◽  
...  
Keyword(s):  
Gall Bladder ◽  
Normal Liver ◽  
Fibroblast Cell ◽  
Coat Pattern ◽  
Liver Structure ◽  
Old Donor ◽  
Cloned Dog ◽  
Original Cell ◽  
Ectopic Liver

The risk of reprogramming abnormalities such as placental hyperdevelopment, excessive fetal growth, or abnormalities of the immune system in cloned neonates is one of the major concerns in cloning research. However, until now, relatively few studies about birth defects have been reported in dog cloning, which might be due to the short in vitro manipulated procedure in this species. Here, we report a cloned dog having abnormal liver and investigated whether the abnormal liver was due to genetic modification. A cloned beagle was produced from a fibroblast derived from a 10-year-old donor, but accidentally died due to cannibalism of a nanny dog on the day of birth. During autopsy, an abnormal liver structure was found; 7 lobes were presented at the normal liver position inside the abdomen, but there was no gall bladder. Interestingly, 3 additional lobes with a gall bladder were found in between the rib and the skin. There were no other macroscopic anomalies observed in this puppy. To evaluate the heredity of this liver abnormality, the liver structure of the donor dog was diagnosed by computed tomography (CT). Also, to assess the possibility of restoring the liver abnormality, recloning was performed using a fibroblast cell line established from the dead pup, and liver positions in the recloned dogs were diagnosed by CT after puberty. In results, 2 recipients delivered 5 recloned dogs with birthweights of 510, 250, 460, 400, and 410 g. The smallest one showed severe bow-legged phenotype in its hind leg, and a unique coat pattern that showed the largest white coat surface. The pup died 10 days after birth, and no other abnormal phenotype was found during autopsy. The other 4 pups showed normal morphology at birth. The CT results showed normal positioning of the liver and gall bladder in all experimental dogs, including the original cell donor dog and recloned dogs. To our knowledge, there have been no reported cases of an ectopic liver and gall bladder present between rib and skin in both cloned and noncloned animals, and we consider that these abnormalities are not a due to genetic cause. Further studies regarding aberrant epigenetic reprogramming in the abnormal liver formation are needed.

73 IMPROVEMENT OF CANINE CLONING EFFICIENCY BY OPTIMIZED DONOR CELL PREPARATION

2010 ◽  
Vol 22 (1) ◽  
pp. 195
Author(s):  
S. W. Park ◽  
Y. W. Jeong ◽  
J. J. Kim ◽  
K. H. Ko ◽  
S. H. Jeong ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Electrical Stimulation ◽  
Somatic Cell ◽  
Somatic Cells ◽  
Contact Inhibition ◽  
Abnormal Development ◽  
Serum Starvation ◽  
Cell Cycle Synchronization ◽  
Donor Cells ◽  
Cloned Dog

The Tibetan Mastiff is the oldest dog breed in the world, and it is at the edge of extinction. Li et al. (2008) believe that protection of and research on the Tibetan Mastiff is extremely urgent, yet few studies have been carried out, particularly at the molecular level. Somatic cell nuclear transfer (SCNT) is an efficient technique for the conservation of endangered animals because it can increase the number of individuals within a population. Considering the virtually unlimited value of cloned canids in critical biotechnology applications, including gene conservation of endangered canids and disease models, the effect of cell-cycle synchronization methods, including the use of cycling canine adult skin fibroblasts (CASF), on the cell-cycle stage and viability of donor nuclei was analyzed. To improve the efficiency of cloned dog production, optimal conditions of donor cells were analyzed by culture duration (Days 1, 2, 3, and 4), passages (2, 4, 7, 10, and 11 passages) and mitotic regulator Plk-1/-4 gene expression. Simerly et al. (2003) reported that the depletion of microtubule motors and centrosomal proteins during enucleation of SCNT procedures caused abnormal development of SCNT embryos. We therefore analyzed Plk-1/-4-induced centriole biogenesis in CASF at different passages of donor cells. In this study, somatic cells were collected from a purebred 9-month-old male Mastiff and an 11-month-old female mastiff. In vivo-matured oocytes were retrieved from outbreed dogs by operation. Cycling cells cultured at Day 4 showed a similar effect to that of cells that were artificially synchronized (contact inhibition or serum starvation). It was also confirmed that fresh and short-term culture (<5 passages) resulted in fewer harmful effects and the same cell viability as control cells, using proliferation assays and expression levels of Plk-1/-4 genes. Therefore, 4 passage-cycling cells at Day 4 were used as donor cells of SCNT. A total of 289 oocytes were reconstructed with each male or female somatic cell and then simultaneously fused/activated with 2 DC pulses of 1.9 kV cm-1 for 30 s of electrical stimulation. Finally, 224 embryos were transferred to 16 naturally synchronized recipients. As a result, we were able to use somatic cells collected from both female and male Tibetan Mastiffs to produce 10 female and 6 male mastiffs. Moreover, one surrogate delivered a quartet of identical cloned female Tibetan Mastiffs puppies; each of 3 surrogates also delivered triplets. Microsatellite analysis demonstrated the genotypic identity of the cloned puppies. In conclusion, the present study shows that (1) cell-cycle synchronization of donor cells by serum starvation/contact inhibition is not required, (2) Plk-1/-4 mRNA can be used to select the donor cells, (3) electrical stimulation alone is sufficient for the activation of SCNT embryos for the production of SCNT cloned dogs, and (4) the cloned dog delivery efficiency (7.1%) was threefold higher than in previous reports. SWP and YWJ contributed equally to this work. WSH was corresponding author and SHH was co-corresponding author.

37 NORMALITY OF NEONATAL REFLEX IN CLONED DOGS

2017 ◽  
Vol 29 (1) ◽  
pp. 126
Author(s):  
E. M. N. Setyawan ◽  
G. A. Kim ◽  
H. J. Oh ◽  
M. J. Kim ◽  
A. Taweechaipaisankul ◽  
...  
Keyword(s):  
Genetic Information ◽  
Nuclear Transfer ◽  
Descriptive Analysis ◽  
Veterinary Science ◽  
Similar Time ◽  
Withdrawal Reflex ◽  
Cloned Dog ◽  
Observed Score ◽  
Phenotypic Instability ◽  
And Training

Since the birth of the world’s first cloned dog, Snuppy, somatic cell nuclear transfer (SCNT) has been a useful tool to propagate the dogs with identical genetic information. However, it is known that cloned animals sometimes exhibit phenotypic instability or abnormality. There have been few investigations about the normality of the neonatal reflex in cloned animals. Therefore, the objective of this study was to evaluate the neonatal reflex in 3 breeds of cloned dogs including shepherd, retriever, and beagle from birth to 28 days of age. Through SCNT, 8 cloned dogs were produced. After birth, 3 types of neonatal reflexes were examined and scored. For examining the flexor dominance reflex, neonatal cloned dogs were held upright and the flexor position of the limb was scored. To evaluate the withdrawal and crossed extensor reflexes, neonates were placed in lateral recumbence and their forelimbs were allowed to relax. Then, the distal forelimbs were pinched and responses were scored according to the frequency and intensity (strong = score 2, variable = score 1, and absent = score 0). The standard responses of neonates were referred from Lindsay et al. (2000 Handbook of Applied Dog Behavior and Training 1, 31–47). Descriptive analysis was used, which was based on the scores from 3 referees who evaluated the videos. The flexor dominance reflex could not be observed (score 0.0) in shepherd by Day 8, in beagle by Day 14 and in retriever by Day 16. Withdrawal reflex began to decrease on Day 22 with score 1.8 for beagle and retriever but decreased in shepherd starting on Day 24 with score 1.8. Crossed extensor reflex for shepherd started to disappear on Day 14 with score 1.5 and completely disappeared (score 0.0) on Day 22; for beagle started to disappear on Day 16 with score 1.8 and was still found until Day 28 with score 1.1; for retriever started to disappear on Day 20 and 28 with score 1.7 and 0.7, respectively. Flexor dominance reflex disappeared in cloned shepherd at a similar time to standard but beagle and retriever seem delayed 6 to 8 days compared with the reference. Withdrawal reflex in all breeds showed normal changes that should persist until adulthood. Cross extensor reflex in shepherd was close to reference but in beagle and retriever was delayed beyond Day 28; this reflex should disappear before adulthood. This study demonstrated that normal neonatal reflexes were identified in the cloned dogs, with some variations among breed. To adapt neonatal reflex as a marker to confirm phenotypic normality in cloned dogs, further investigation using various breeds of cloned dogs and greater numbers of subjects is needed. This study was supported by IPET (#316002-05-1-SB010), RDA (#PJ010928032016), Research Institute for Veterinary Science, Natural Balance Korea and the BK21 plus program.

Pharmacological characterisation of a cloned dog 5-HT1B receptor cell line

1998 ◽  
Vol 360 (1) ◽  
pp. 117-121 ◽  
Author(s):  
Margaret S. Beer ◽  
M.Anne Heald ◽  
George McAllister ◽  
Josephine A. Stanton
Keyword(s):  
Cell Line ◽  
Receptor Cell ◽  
Cloned Dog

285 GENERATION OF CANINE INDUCED PLURIPOTENT STEM CELLS FROM CANINE FETAL FIBROBLAST AND ADULT FIBROBLAST OF CLONED DOG

2013 ◽  
Vol 25 (1) ◽  
pp. 290 ◽  
Author(s):  
H. S. Kwon ◽  
H. J. Oh ◽  
D. H. Lee ◽  
D. E. Kim ◽  
S. K. Kang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
In Vitro Differentiation ◽  
Germ Layers ◽  
Fetal Fibroblast ◽  
Reprogramming Factors ◽  
Induced Pluripotent ◽  
Cloned Dog

Induced pluripotent stem cells (iPSC) derived from a patient’s fibroblasts have been used as fine resources for studying disease mechanisms and therapeutic strategies. The dog is considered invaluable in human disease research because its genetic diseases are strikingly similar to those of human. Therefore, we generated cloned dogs and transgenic cloned dogs via somatic cell nuclear transfer. In this study, we tried to derive canine iPSCs from canine fibroblasts to establish a way to make iPSC from skin fibroblasts of transgenic cloned dogs. We isolated canine fetal fibroblast (FF) from normal beagles and adult skin fibroblast (ASF) from cloned beagles. Both ASF and FF were infected with all-in-one retroviral vector that delivers human reprogramming factors (Oct4, Sox2, Klf4, c-Myc). Ten to twenty-one days after infection, the colony-shaped structure was picked and plated on a mouse embryonic fibroblast (MEF) feeder layer, pretreated with mitomycin C. Then, all cells were cultured with DMEM/F12 supplemented with 20% fetal bovine serum, 5 ng mL–1 basic fibroblast growth factor (bFGF), 5 ng mL–1 LIF, 0.1 mM β-mercaptoethanol, 1% NEAA, and 1% penicillin-streptomycin. Alkaline phosphatase (AP) activity and expression of Oct4, Sox2, SSEA1, and SSEA4, were observed in the cells to characterise the iPS cell colonies. In vitro differentiation of 10th-passage canine iPSC was performed through embryonic body formation. About 50 canine iPS-like colonies were formed on a 100-mm dish. As a result, the canine iPSC from FF (iPSC-FF) and canine iPSC from ASF (iPSC-ASF) showed typical colony morphology, and both stained positively for AP. The expression of pluripotency-associated transcription factors Oct4 and Sox2 was positively displayed in iPSC-FF colonies. The stem cell markers SSEA1 and SSEA4 were negative in canine iPSC-FF. The canine iPS-FF spontaneously differentiated into all 3 germ layers in vitro, showing positive expressions of βIII-tubulin (ectoderm), α-SMA (mesoderm), and GATA6 (endoderm). As for iPS-ASF, characterisation and in vitro differentiation experiment are in progress. These results show that canine iPS-FF are similar to embryonic stem cells in terms of morphology and the ability to differentiate into 3 germ layers. Although we did not demonstrate complete verification of canine iPS-ASF of the cloned dog, their morphology, AP expression, and iPS-FF generation should indicate the possibility of iPSC production in the cloned dog. In conclusion, retroviral transduction of 4 human reprogramming factors can reprogram canine fetal fibroblasts into canine iPSC. The technique of producing canine iPSC will stimulate the utilisation of transgenic cloned dogs and expand the range of human diseases or therapeutic application. This study was supported by RDA (#PJ0089752012), RNL Bio (#550-20120006), IPET (#311011-05-1-SB010), Research Institute for Veterinary Science, and Nestlé Purina Korea.

First cloned dog created in Korea

2005 ◽  
Vol 157 (7) ◽  
pp. 183-183
Keyword(s):  
Cloned Dog

Ectopic liver and gallbladder in a cloned dog: Possible nonheritable anomaly

2015 ◽  
Vol 84 (6) ◽  
pp. 995-1002 ◽  
Author(s):  
Min Jung Kim ◽  
Sang Chul Kang ◽  
Jae Hwan Kim ◽  
Hyun Ju Oh ◽  
Geon A Kim ◽  
...  
Keyword(s):  
Cloned Dog ◽  
Ectopic Liver

Employing mated females as recipients for transfer of cloned dog embryos

2013 ◽  
Vol 25 (4) ◽  
pp. 700 ◽  
Author(s):  
Geon A Kim ◽  
Hyun Ju Oh ◽  
Jung Eun Park ◽  
Min Jung Kim ◽  
Eun Jung Park ◽  
...  
Keyword(s):  
Caesarean Section ◽  
Somatic Cell Nuclear Transfer ◽  
Nuclear Transfer ◽  
Progesterone Concentration ◽  
Due Date ◽  
Cloned Dog ◽  
First Time ◽  
Implantation And Pregnancy Rates ◽  
Parthenogenetic Embryos

It has been suggested that co-transferring parthenogenetic embryos could improve the pregnancy success rate with cloned embryos in mammals. As an alternative to co-transferring parthenotes, in dogs we employed recipient females that possessed in vivo-fertilised embryos as a result of mating to determine whether mated bitches could be suitable recipients for cloned embryos. The effect of using mated recipients on implantation and pregnancy rates of canine somatic cell nuclear transfer embryos was also determined. Cloned embryos were transferred into the oviducts of naturally synchronous females that had mated with male dogs before ovulation. The pregnancy rate appeared to be similar between mated recipients (50%) and non-mated recipients (28.57%; P > 0.05). However, the delivery rate of cloned pups was significantly higher in mated recipients than non-mated recipients (10.53 vs 2.38%; P < 0.05). A decrease in progesterone levels in the mated recipients before the due date induced natural delivery. However, cloned pups in non-mated recipients were delivered by Caesarean section because the fall in progesterone concentration in these females did not occur until the due date. The present study demonstrated for the first time that mated female dogs can be used as recipients for cloned embryos.

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