34 MEIOTIC RECOMBINATION IN SOMATIC CELL NUCLEAR TRANSFER BULLS AND THEIR OFFSPRING

2008 ◽  
Vol 20 (1) ◽  
pp. 97
Author(s):  
E. J. Hart ◽  
A. Pinton ◽  
A. Powell ◽  
R. Wall ◽  
W. A. King
Keyword(s):  
Somatic Cell ◽  
Synaptonemal Complex ◽  
Meiotic Recombination ◽  
Somatic Cell Nuclear Transfer ◽  
Nuclear Transfer ◽  
Mixed Model ◽  
Genetic Material ◽  
Repeated Measurements ◽  
Meiotic Process ◽  
Mixed Model Analysis

In mammals, homologous chromosome pairing and recombination are essential events for meiosis. The generation of reciprocal exchanges of genetic material ensure both genetic diversity and the proper segregation of homologous chromosomes. With the advent of reproductive biotechnologies such as somatic cell nuclear transfer (SCNT) in the livestock industry, these vital steps have begun to be bypassed. Despite this, there have been few studies carried out on the reproductive characteristics of cloned animals and none to date regarding the consequences on the meiotic process. As these procedures grow in popularity and use, the importance of evaluating the long-term viability, health, and productivity of cloned animals and any subsequent offspring in future generations to validate potential applications of the technology is increasingly evident. Previously, studies of recombination and synapsis have focused on the physical observation of chiasmata formation in meiotic chromosomes; however, in recent years, the characterization of proteins that localize to the sites of crossing-over and of proteins present in the synaptonemal complex have permitted the study of meiotic recombination using a precise direct immunocytogenetic approach. Cytological analysis of meiotic cells obtained from the testicular tissue of five normal bulls of proven fertility, two SCNT transgenic bulls (Powell et al. 2004 Biol. Reprod. 71, 210–216, 2004), and four reproductively mature offspring of SCNT bulls was performed in order to detect the effects of SCNT on the meiotic process. Over 50 pachytene cells per animal were analyzed by immunofluorescence using antibodies against the synaptonemal complex protein 3 (SCP3) and the mismatch repair protein 1 (MLH1) located on the crossover sites. Data were analyzed using a mixed model analysis of variance with repeated measurements to determine group effects (SAS 9.1; SAS Institute, Inc., Cary, NC, USA). The average number of crossovers per spermatocyte for the non-SCNT bulls (42 � 4 SD, min: 33, max: 56), SCNT bulls (43 � 5 SD, min: 35, max: 56), and SCNT offspring (46 � 4 SD, min: 37, max: 58) were quite similar among the cells of the same individual; however, inter-individual variation was observed. These results are the first documentation of the normal range of variability of recombination distribution within the cattle genome and suggest that the SCNT process does not affect meiotic recombination. This work was funded by NSERC and the CRC program.

32 CLONING OF ELITE QUARANTINE SNIFFING DOG BY SOMATIC CELL NUCLEAR TRANSFER

2013 ◽  
Vol 25 (1) ◽  
pp. 164
Author(s):  
H. J. Oh ◽  
M. J. Kim ◽  
G. A. Kim ◽  
J. Choi ◽  
E. J. Park ◽  
...  
Keyword(s):  
Somatic Cell ◽  
Somatic Cell Nuclear Transfer ◽  
Nuclear Transfer ◽  
Mixed Model ◽  
Linear Mixed Model ◽  
Assisted Reproductive Technologies ◽  
Reproductive Technologies ◽  
Donor Cells ◽  
Wide Range ◽  
Sound Sensitivity

Somatic cell nuclear transfer (SCNT) in assisted reproductive technologies has been considered for the conservation of valuable or endangered animals. Dogs that were originally bred for hunting, such as beagles, have an exceptional ability to detect a particular smell from many others. For that reason, the beagles have been used to detect quarantine risk items from a wide range of goods in assorted luggage without scaring or disrupting the passengers. Though very useful and highly in need, elite quarantine sniffing beagles with excellent abilities are rare; much time, effort, and money are required in producing them. Here, we have applied SCNT for propagation of elite quarantine sniffing dogs to save time and economic burden. Ear fibroblasts from a 10-year-old adult male elite quarantine sniffing beagle were isolated and cultured in vitro as donor cells. For SCNT, in vivo-matured oocytes, obtained by flushing the uterine tubes of oocyte donors (mixed breed), were used. The oocytes were enucleated, microinjected with donor cells, fused by electrical stimulation, and activated chemically. Reconstructed oocytes were surgically transferred into the uterine tube of naturally synchronous recipient females. A total of 212 activated cloned embryos were transferred into 12 female recipient dogs and 4 recipients became pregnant. The 4 pregnant recipients delivered 4 pups through caesarean section or natural delivery, but 1 died right after birth and did not show an abnormality. Other live puppies exhibited normal phenotypes; their appearance was similar to that of the donor dog. All cloned pups were genetically identical to the donor dog and their mitochondrial DNA was from their oocyte donor dogs. When the cloned pups were 16 weeks old, we conducted a Volhard test, which is commonly used to describe the following puppy aptitudes: social attraction, following, restraint, social dominance, elevation dominance, retrieving, touch sensitivity, sound sensitivity, and sight sensitivity. Dog behavior data on differences in transcript abundance were analyzed by a general linear mixed model. The 3 cloned pups showed similar behavioral tendencies. The present study demonstrates that NT technique using donor cell derived from 1 elite quarantine sniffing dog is useful to produce a large number of quarantine sniffing dogs. This study was supported by RDA (no. PJ0089752012), RNL Bio (no. 550-20120006), IPET (no. 311062-04-1-SB010), Research Institute for Veterinary Science, Nestlé Purina Korea, and TS Corporation.

Advances in equine cloning by somatic cell nuclear transfer (SCNT) technique in horses: a review

2016 ◽  
Vol 5 (12) ◽  
pp. 5124
Author(s):  
Rajesh Wakchaure ◽  
Subha Ganguly*
Keyword(s):  
In Vitro Culture ◽  
Somatic Cell ◽  
Oocyte Maturation ◽  
Somatic Cell Nuclear Transfer ◽  
Nuclear Transfer ◽  
Genetic Material ◽  
Culture Conditions ◽  
Preimplantation Embryos ◽  
Oocyte Activation

Cloning a horse means using the genetic material (DNA) from a donor horse to produce a genetically identical foal. This technique involves collecting the DNA from the donor and inserting that DNA into an egg from another mare whose, DNA content has been removed, fusing donor nucleus with enucleated recipient oocytes, which then develops as an embryo, in vitro culture of embryo and lastly transfer cultured embryo into the uterus of a recipient mare. The modification of the in vitro culture conditions which can be suitable for equine oocyte activation, oocyte maturation and embryo development are the fundamental steps for a successful in vitro procedure for somatic cell nuclear transfer (SCNT) in the horse to avoid the embryo losses. In general, few studies are available in the literature on equine in vitro embryo production and it is only recently that reports have been published on completely in vitro production of equine preimplantation embryos by means of in vitro oocyte maturation The present review discusses the latest developments in the field of equine cloning technique with the employment of SCNT. The basic understanding of SCNT for in vitro culture conditions is relevant to the increased efficiency of cloning. The available genotype can be used by SCNT which can enhance the vigour of a particular infertile or low fertile animal to produce normal fertility.

scholarly journals Author Correction: Application of interspecific Somatic Cell Nuclear Transfer (iSCNT) in sturgeons and an unexpectedly produced gynogenetic sterlet with homozygous quadruple haploid

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Effrosyni Fatira ◽  
Miloš Havelka ◽  
Catherine Labbé ◽  
Alexandra Depincé ◽  
Viktoriia Iegorova ◽  
...  
Keyword(s):  
Somatic Cell ◽  
Somatic Cell Nuclear Transfer ◽  
Nuclear Transfer

scholarly journals Stem cell therapies and benefaction of somatic cell nuclear transfer cloning in COVID-19 era

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Birbal Singh ◽  
Gorakh Mal ◽  
Vinod Verma ◽  
Ruchi Tiwari ◽  
Muhammad Imran Khan ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Regenerative Medicine ◽  
Somatic Cell ◽  
Somatic Cell Nuclear Transfer ◽  
Nuclear Transfer ◽  
Drug Testing ◽  
Human Interaction ◽  
Patient Specific ◽  
Wide Range

Abstract Background The global health emergency of COVID-19 has necessitated the development of multiple therapeutic modalities including vaccinations, antivirals, anti-inflammatory, and cytoimmunotherapies, etc. COVID-19 patients suffer from damage to various organs and vascular structures, so they present multiple health crises. Mesenchymal stem cells (MSCs) are of interest to treat acute respiratory distress syndrome (ARDS) caused by SARS-CoV-2 infection. Main body Stem cell-based therapies have been verified for prospective benefits in copious preclinical and clinical studies. MSCs confer potential benefits to develop various cell types and organoids for studying virus-human interaction, drug testing, regenerative medicine, and immunomodulatory effects in COVID-19 patients. Apart from paving the ways to augment stem cell research and therapies, somatic cell nuclear transfer (SCNT) holds unique ability for a wide range of health applications such as patient-specific or isogenic cells for regenerative medicine and breeding transgenic animals for biomedical applications. Being a potent cell genome-reprogramming tool, the SCNT has increased prominence of recombinant therapeutics and cellular medicine in the current era of COVID-19. As SCNT is used to generate patient-specific stem cells, it avoids dependence on embryos to obtain stem cells. Conclusions The nuclear transfer cloning, being an ideal tool to generate cloned embryos, and the embryonic stem cells will boost drug testing and cellular medicine in COVID-19.

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