80 USE OF THE SHORT DISPOSABLE NEEDLE SYSTEM FOR FOLLICLE ASPIRATION IN MARES

2010 ◽  
Vol 22 (1) ◽  
pp. 198
Author(s):  
K. Song ◽  
W. Lee ◽  
Y. Chun ◽  
I. Lee ◽  
S. Yeon ◽  
...  
Keyword(s):  
Stainless Steel ◽  
Embryo Transfer ◽  
Recovery Rate ◽  
Nuclear Transfer ◽  
Double Lumen ◽  
Stainless Steel Needle ◽  
Disposable Needle ◽  
Follicle Aspiration

To achieve success in equine somatic cell nuclear transfer (SCNT), it is important to obtain recipient oocytes of good quality. Transvaginal ultrasoundguided follicle aspiration (TVUFA) is one of the methods to obtain recipient oocytes in equine SCNT, but the commercial long double-lumen needle for TVUFA in large animals is not currently purchasable. The aims of the present study were (1) to compare the recovery rate of short disposable needle system (14G) with that of the long double-lumen needle (12G) and (2) to investigate the developmental competency of recovered oocytes after SCNT and embryo transfer (ET). A real-time ultrasound scanner (Mylab30 vet, Esaote, Italy) equipped with a 7.5-MHz convex array transducer (model EC123) housed in a plastic vaginal device with stainless steel needle guidance was used for TVUFA from pre-ovulatory follicles 25 to 40 mm in diameter on synchronized thoroughbred mares between 6 and 9 years of age. Two types of needles were used: (1) a 12G double-lumen long needle (V-EOAD-1260L; Cook, Brisbane, Australia); (2) a 14G single-lumen disposable needle (2.1 × 80mm; Bovi-vet, Kruuse, Denmark) inserted with 18G inner needle (0.8 × 600 mm) using stainless steel connector and tube. Recovered oocytes were matured in vitro in TCM-199 with 5 mU mL-1 FSH (Folltropin-V, Bioniche, Belleville, ON, Canada) and 10% fetal bovine serum (Sigma, St. Louis, MO, USA) for 12 to 16 h. Matured oocytes were enucleated and electrically fused with equine skin fibroblasts (2.25 kV cm-1, 20μs, 2 pulses). Fused couplets were activated with 5 μM ionomycin for 4 min followed by 5 h culture in 1 mM 6-DMAP. Immediately following SCNT procedures, cloned embryos were surgically transferred to the oviducts of recipient mares (n = 2 to 5 embryos per recipient) that had ovulated within 24 hours before the transfer. An initial pregnancy examination was performed using transrectal ultrasonography between Days 14 and 16 (Day 0 = surgery). The recovery rate of the short disposable needle (n = 89, 44.1%) was slightly increased compared with that of the long needle (n = 34, 29.8%), but the difference was not significant. Nineteen SCNT embryos were transferred to 8 mares, and 1 mare is maintaining pregnancy for 60 days. The results of this study demonstrated that our short disposable needle system could be used instead of the commercial long needle and in vivo development of oocyte recovered with the 14G needle could be maintained after nuclear transfer and embryo transfer.

scholarly journals 60 HISTOLOGICAL COMPARISONS BETWEEN NUCLEAR TRANSFER AND IN VIVO PORCINE EMBRYOS

2005 ◽  
Vol 17 (2) ◽  
pp. 180
Author(s):  
L. Overman ◽  
L. Lai ◽  
H.-T. Cheong ◽  
G.-S. Im ◽  
K.-W. Park ◽  
...  
Keyword(s):  
Embryo Transfer ◽  
Nuclear Transfer ◽  
High Rate ◽  
Normal Numbers ◽  
Diagnostic Laboratory ◽  
Invaluable Tool ◽  
Embryonic Loss ◽  
Mitotic Figures

Due to the high rate of embryonic loss during a nuclear transfer pregnancy, cloning is considered a relatively inefficient process. However, as the only method of producing knockout domestic animals it is considered an invaluable tool for the biotechnical industry. By histologically comparing embryos at significant stages in the porcine pregnancy (Days 10, 12, and 14), factors contributing to embryonic loss may be revealed. Many consider the period between days 10 and 14 to be critical for determining survivability as the embryos must undergo rapid changes to signal for maternal recognition of pregnancy as well as adapt to a changing environment. This study included three gilts per stage of pregnancy and four different experimental groups for each stage studied: nonpregnant animals, in vivo-pregnant animals, nuclear transfer (NT) recipients, and in vitro-manipulated recipients (IVM). IVM embryos were in vitro-produced embryos upon which a mock nuclear transfer has been performed in an effort to account for the variability introduced by the actual technique. Animals either were bred or underwent a surgical embryo transfer on Day 1 of the estrous cycle according to their assigned experimental group. Fifty embryos were transferred per embryo transfer. Embryos were flushed from the uterine horns at time of collection (Day 10, 12, or 14) and preserved in 10% neutral buffered formalin. All embryonic disc diameters and gross morphology were evaluated as parameters for normal development. Embryos were then dehydrated in ethanol, paraffin-imbedded, sectioned, stained with hemotoxylin and eosin, and Day 14 embryos were evaluated for abnormalities such as higher-than-normal nucleoli numbers, increased cytoplasmic vacuoles, and higher than normal numbers of mitotic figures. All results were analyzed using ANOVA. There were significant differences (P < .0001) between diameters of the embryonic disks, with the diameters of the NT embryonic disks being smaller than those of the in vivo controls at all stages studied. Morphologically, the in vivo controls were more developmentally competent than their NT counterparts by the time they reached Day 14 (P = 0.0002) in that most had achieved the more advanced elongated form of growth as opposed to remaining spherical in shape. Significant histological differences in the number of nucleoli per nuclei were also found between in vivo and NT embryos (P = 0.05) as well as between MNT and NT embryos (P = 0.05). Therefore, nuclear transfer embryos develop at a much slower rate than their in vivo counterparts and often exhibit histological abnormalities that could contribute to this slow growth. Due to the apparent increase in nucleoli, it is possible that NT embryos are being arrested at a specific stage in the cell cycle. The authors would like to acknowledge the Research Animal Diagnostic Laboratory for their help in imbedding, sectioning, and staining the embryos; Dr. Duane Keisler for running the hormone assays; and Kristin Whitworth, Melissa Samuel, Aaron Bonk, Jin-Geol Kim, Emily Fergason, David Wax, Tom Cantley, August Rieke, Randy Farrell, and Lacey Griesbaum for all their help.

ECONOMIC PERSPECTIVES OF BIOTECHNOLOGIES USING IN ANIMAL FARMING

1970 ◽  
pp. 57-60
Author(s):  
I. F. Gorlov ◽  
A. A. Mosolov ◽  
G. V. Komlatskiy ◽  
M. A. Nesterenko ◽  
K. D. Nimbona ◽  
...  
Keyword(s):  
Embryo Transfer ◽  
Economic Effect ◽  
Dairy Herd ◽  
State Level ◽  
Modern Level ◽  
Economic Perspectives ◽  
The Cost ◽  
Short Time

The article presents materials on the study of the possibility of reproduction and increase in the herd of highly productive cows through the use of embryo transplantation technology. The classical (in vivo) and more modern, developing (in vitro) methods of embryotransfer, their positive and negative sides are considered in detail. The possibility of accelerating the breeding process by using the method of transplantation, in which from one cow can be obtained from 10 to 100 calves, which will allow for 4-5 years, almost any herd (of any size and breed) with the help of biotechnology to turn into a cattle-breeding enterprise of the most modern level. At the same time, heifers obtained from unproductive cows can be used as "surrogate" mothers who are transplanted with the best donor embryos, which allows to obtain a full-fledged offspring adapted to local environmental conditions. A detailed scheme of obtaining, evaluation, storage, as well as the cost and economic effect of embryo transplantation was calculated, the market was evaluated, the required annual volume of transplants and the number of donor cows for large livestock farms were determined. As a positive example of "Scientific-production enterprise "Centre of biotechnology and embryo transfer" in 2014, implemented a project for accelerated replacement and genetic improvement of the dairy herd, engraftment averaged 57-69%, and the economic effect of the enterprise from getting a single animal by the method of embryo transfer, compared with imports of similar close in quality, ranged from 60 to 100 thousand rubles on his head. It is shown that it is necessary to organize at the state level a developed service for embryo transplantation to reduce the cost of embryo transfer and the possibility of creating in a short time in the country's own highly productive breeding nucleus of dairy and beef cattle, which will reduce, and in the future completely eliminate, import dependence on cattle products.

Effects of rate of development of in vitro-produced ovine embryos on sex ratio and in vivo survival after embryo transfer

1997 ◽  
Vol 48 (8) ◽  
pp. 1369-1378 ◽  
Author(s):  
S.L. Catt ◽  
J.K. O'Brien ◽  
W.M.C. Maxwell ◽  
G. Evans
Keyword(s):  
Sex Ratio ◽  
Embryo Transfer ◽  
Rate Of Development

scholarly journals 64COMPARISON OF DEVELOPMENTAL POTENTIAL OF IN VIVO AND IN VITRO RECIPIENT OOCYTES AFTER NUCLEAR TRANSFER IN GOAT

2004 ◽  
Vol 16 (2) ◽  
pp. 154
Author(s):  
H.S. Park ◽  
M.Y. Lee ◽  
S.P. Hong ◽  
J.I. Jin ◽  
J.K. Park ◽  
...  
Keyword(s):  
Nuclear Transfer ◽  
Electric Pulse ◽  
Serum Starvation ◽  
Cleavage Rate ◽  
Developmental Potential ◽  
In Vitro Development ◽  
Significant Difference ◽  
Vitro Development

Recent techniques in somatic cell nuclear transfer (SCNT) have been widely used for animal research. In addition, SCNT techniques may allow for the rescue of endangered species. Despite efforts for wildlife preservation, however, some threatened or endangered wild animal species will likely become extinct. As a preliminary experiment of a series in wildlife research, we tried to identify an improved method for the production of more transferable NT embryos in goats. Mature donor animals of Korean native goats (20–25kg) were synchronized with a CIDR (type G; InterAg, New Zealand) vaginal implant for 10 days followed by a total of 8 twice daily injections of 70mg of FSH (Folltropine, London, Ontario, Canada) and 400IU of hCG (Chorulon, Intervet, Moxmeer, The Netherlands). Oocytes were then collected surgically by retograde oviduct flush or direct aspiration from ovarian follicles in vivo at 29–34h after hCG. Oocytes collected from follicles were matured in TCM-199 containing 10% FBS and hormones. Prepared ear skin cells from the goat were cultured in TCM-199 containing 10% FBS at 39°C, 5% CO2 in air, and confluent monolayers were obtained. Oocytes were enucleated and donor cells from serum starvation (0.5%) culture were fused through a single electric pulse (DC 2.36kvcm−1, 17μs), and then activated by a single electric pulse (AC 5vmm−1, 5s+DC 1.56kvcm−1, 30μs) or chemical treatment (5μgmL−1 ionomycin 5min−1, 1.9mM 6-DMAP/4h). Reconstructed oocytes were cultured in M16 medium with 10% goat serum (GS) for 6–7 days. Data were analyzed by chi-square test. In in vitro development, significantly (P&lt;0.05) more oocytes were cleaved (24/30, 80.0%) and developed (7/24, 29.2%) to morula or blastocyst stage, respectively, in NT oocytes activated by Iono + DMAP compared to electric stimulated oocytes (2/21, 40.0%; 0/2, 0%). There was a significant difference in in vitro development of NT embryos by the method of oocyte collection. Cleavage rate was higher (P&lt;0.05) in NT embryos from in vivo oocytes (23/28, 82.1%) than in in vitro matured oocytes (19/35, 54.3%), and further development to morula or blastocyst was also significantly (P&lt;0.05%) higher in NT embryos from in vivo oocytes (7/23, 30.4%) than in NT embryos from in vitro matured oocytes (0/19, 0%). When we compared NT embryos to parthenotes, developmental rate was not significantly different between NT embryos and parthenotes. These results strongly suggest that the in vivo oocytes will have superior developmental potential to oocytes matured in vitro. Table 1 Effect of different oocyte source on in vitro development following caprine SCNT

scholarly journals Comparative analysis of different nuclear transfer techniques to prevent the transmission of mitochondrial DNA variants

2019 ◽  
Author(s):  
M Tang ◽  
R R Guggilla ◽  
Y Gansemans ◽  
M Van der Jeught ◽  
A Boel ◽  
...  
Keyword(s):  
Mitochondrial Dna ◽  
Disease Transmission ◽  
Nuclear Transfer ◽  
Smooth Endoplasmic Reticulum ◽  
Scientific Evidence ◽  
Polar Body ◽  
Comparable Rate ◽  
Carry Over

Abstract Prevention of mitochondrial DNA (mtDNA) diseases may currently be possible using germline nuclear transfer (NT). However, scientific evidence to compare efficiency of different NT techniques to overcome mtDNA diseases is lacking. Here, we performed four types of NT, including first or second polar body transfer (PB1/2T), maternal spindle transfer (ST) and pronuclear transfer (PNT), using NZB/OlaHsd and B6D2F1 mouse models. Embryo development was assessed following NT and mtDNA carry-over levels were measured by next generation sequencing (NGS). Moreover, we explored two novel protocols (PB2T-a and PB2T-b) to optimize PB2T using mouse and human oocytes. Chromosomal profiles of NT-generated blastocysts were evaluated using NGS. In mouse, our findings reveal that only PB2T-b successfully leads to blastocysts. There were comparable blastocyst rates amongst PB1T, PB2T-b, ST and PNT embryos. Furthermore, PB1T and PB2T-b had lower mtDNA carry-over levels than ST and PNT. After extrapolation of novel PB2T-b to human in vitro matured (IVM) oocytes and in vivo matured oocytes with smooth endoplasmic reticulum aggregates (SERa) oocytes, the reconstituted embryos successfully developed to blastocysts at a comparable rate to ICSI controls. PB2T-b embryos generated from IVM oocytes showed a similar euploidy rate to ICSI controls. Nevertheless, our mouse model with non-mutated mtDNAs is different from a mixture of pathogenic and non-pathogenic mtDNAs in a human scenario. Novel PB2T-b requires further optimization to improve blastocyst rates in human. Although more work is required to elucidate efficiency and safety of NT, our study suggests that PBT may have the potential to prevent mtDNA disease transmission.

scholarly journals In vivo culture of bovine embryos and quality assessment of in vivo vs. in vitro produced embryos

2012 ◽  
Vol 50 (No. 4) ◽  
pp. 149-158 ◽  
Author(s):  
V. Havlicek ◽  
M. Lopatarova ◽  
S. Cech ◽  
R. Dolezel ◽  
T. Huber ◽  
...  
Keyword(s):  
Quality Assessment ◽  
Recovery Rate ◽  
Bovine Embryos ◽  
In Vivo Culture ◽  
Blastocyst Rate ◽  
Culture Procedure ◽  
Recovery Methods

Routine access to the bovine oviduct for in vivo culture accomplishes various demands on embryo production for scientific as well as commercial purposes. The experiments conducted in the present study focused on the efficiency of recovery methods after temporary in vivo culture of bovine embryos in oviducts of the homologous species using transvaginal endoscopy (Experiment I) and on the quality assessment of recovered blastocysts (Experiment II). In Experiment I in vitro matured oocytes were fertilized, cultured for 1 to 3 days and transferred unilaterally into the ipsilateral oviducts of 54 heifers by the means of transvaginal endoscopy. After 4 to 6 days of in vivo culture embryos were re-collected either by non-surgical flushing of uterine horns (U-group) or by combined flushing of the oviducts and uterine horns (OU-group). In total the recovery rate was 38.4% (780/2029). After flushing at day seven, 106 blastocysts (blastocyst rate: 13.6% ) were found. The additional 24 h of in vitro culture (day eight) resulted in 153 blastocysts (blastocyst rate: 19.6% ). The recovery rate in the OU-group was twice as efficient as in the U-group (390/1358 vs. 390/671, P &lt; 0.01). The recovery rates among the different stages of transferred embryos did not differ significantly; likewise cross-effects among the stages and the recovery methods were non-significant. The recovery methods (P &lt; 0.001) and the interaction between the recovery methods and the stages of transferred embryos (P &lt; 0.01) had an influence on blastocyst yields on day seven (U-group 37/1358 vs. OU-group 69/671) and day eight (U-group 48/1358 vs. OU-group 105/671). In Experiment II embryo quality was assessed by the survival rate of blastocysts after freezing in ethylene glycol. Day seven embryos were produced in vitro (in vitro group D7) or by IVM/IVF followed by a combined culture procedure (2 to 3 days in vitro prior to 4&nbsp;to 5 days in vivo) (in vivo group D7) or after superovulation and collection at day seven (superovulation group). Embryos from in vitro group D7 re-expanded only for 6 h after thawing, embryos from in vivo group D7 and superovulation group were alive for 24 h and 72 h of culture, respectively. Only embryos derived by superovulation showed hatching activity. Blastocysts from the in vitro group D7 and the in vivo group D7 that were held in culture medium for additional 24 h (day eight) showed an analogous post-thawing culture behaviour. In conclusion, the results of the present study demonstrated that some embryos transferred for in vivo culture remain in the oviduct even at day seven. Hence, combined flushing of oviducts and uterine horns after in vivo culture in the bovine oviduct is necessary for effective embryo re-collection. The quality of recovered embryos after temporary in vivo culture assessed by cryotolerance was in-between those produced in vitro or recovered after superovulation.

Differential effects of culture and nuclear transfer on relative transcript levels of genes with key roles during preimplantation

Zygote ◽  
2006 ◽  
Vol 14 (1) ◽  
pp. 81-87 ◽  
Author(s):  
P.N. Moreira ◽  
R. Fernández-Gonzalez ◽  
M.A. Ramirez ◽  
M. Pérez-Crespo ◽  
D. Rizos ◽  
...  
Keyword(s):  
Nuclear Transfer ◽  
Culture Medium ◽  
Cumulus Cell ◽  
Blastocyst Stage ◽  
Mrna Levels ◽  
Differential Effects ◽  
Glut 1 ◽  
Mouse Somatic Cell

It is well known that the preimplantation culture environment to which embryos are exposed influences the expression of developmentally important genes. Recently, it has been reported that MEMα, a culture medium commonly used for somatic cells, allows high rates of preimplantation development and development to term of mouse somatic cell nuclear transfer (SCNT) embryos. The objective of this study was to compare the differential effects of this medium and of the nuclear transfer procedure on the relative mRNA abundance of several genes with key roles during preimplantation. The relative mRNA levels of nine genes (Glut 1, Glut 5, G6PDH, Bax, Survivin, Gpx 1, Oct4, mTert and IGF2bp1) were quantified at blastocyst stage on cumulus cell cloned embryos cultured in MEMα, as well as on in vivo cultured and MEMα cultured controls. Only three of the nine transcripts analysed (Glut 5, Gpx 1 and Igf2bp1) were significantly down-regulated at blastocyst stage in in vitro produced controls. However, most genes analysed in our MEMα cultured cloned embryos showed altered transcription levels. Interestingly, between cloned and in vitro produced controls only the transcription levels measured for Glut 1 were significantly different. This result suggests that Glut 1 may be a good marker for embryo quality after cumulus cell nuclear transfer.

scholarly journals Simultaneous gene quantitation of multiple genes in individual bovine nuclear transfer blastocysts

Reproduction ◽  
2007 ◽  
Vol 133 (1) ◽  
pp. 231-242 ◽  
Author(s):  
Craig Smith ◽  
Debbie Berg ◽  
Sue Beaumont ◽  
Neil T Standley ◽  
David N Wells ◽  
...  
Keyword(s):  
Gene Expression ◽  
Nuclear Transfer ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Ectopic Expression ◽  
Cell Number ◽  
Transcript Levels ◽  
Multiple Genes

During somatic cell nuclear transfer (NT), the transcriptional status of the donor cell has to be reprogrammed to reflect that of an embryo. We analysed the accuracy of this process by comparing transcript levels of four developmentally important genes (Oct4,Otx2,Ifitm3,GATA6), a gene involved in epigenetic regulation (Dnmt3a) and three housekeeping genes (β-actin, β-tubulinandGAPDH) in 21 NT blastocysts with that in genetically half-identicalin vitroproduced (IVP,n=19) andin vivo(n=15) bovine embryos. We have optimised an RNA-isolation and SYBR-green-based real-time RT-PCR procedure allowing the reproducible absolute quantification of multiple genes from a single blastocyst. Our data indicated that transcript levels did not differ significantly between stage and grade-matched zona-free NT and IVP embryos except for Ifitm3/Fragilis, which was expressed at twofold higher levels in NT blastocysts.Ifitm3expression is confined to the inner cell mass at day 7 blastocysts and to the epiblast in day 14 embryos. No ectopic expression in the trophectoderm was seen in NT embryos. Gene expression in NTand IVP embryos increased between two- and threefold for all eight genes from early to late blastocyst stages. This increase exceeded the increase in cell number over this time period indicating an increase in transcript number per cell. Embryo quality (morphological grading) was correlated to cell number for NT and IVP embryos with grade 3 blastocysts containing 30% fewer cells. However, only NT embryos displayed a significant reduction in gene expression (50%) with loss of quality. Variability in gene expression levels was not significantly different in NT, IVP orin vivoembryos but differed among genes, suggesting that the stringency of regulation is intrinsic to a gene and not affected by culture or nuclear transfer.Oct4levels exhibited the lowest variability. Analysing the total variability of all eight genes for individual embryos revealed thatin vivoembryos resembled each other much more than did NT and IVP blastocysts. Furthermore,in vivoembryos, consisting of 1.5-fold more cells, generally contained two- to fourfold more transcripts for the eight genes than did their cultured counterparts. Thus, culture conditions (in vivoversusin vitro) have greater effects on gene expression than does nuclear transfer when minimising genetic heterogeneity.

102 RABBIT CLONING: HISTONE ACETYLATION STATUS OF DONOR CELLS AND CLONED EMBRYOS

2007 ◽  
Vol 19 (1) ◽  
pp. 168
Author(s):  
V. Zakhartchenko ◽  
F. Yang ◽  
R. Hao ◽  
E. Wolf
Keyword(s):  
Histone Acetylation ◽  
Nuclear Transfer ◽  
Epigenetic Mark ◽  
Cloning Efficiency ◽  
Developmental Potential ◽  
Acetylation Status ◽  
Donor Cells ◽  
Fetal Fibroblasts

Epigenetic status of the genome of a donor nucleus is likely to be associated with the developmental potential of cloned embryos produced by somatic cell nuclear transfer (SCNT). Prevention of epigenetic errors by manipulation of the epigenetic status of donor cells is expected to result in improvement of cloning efficiency. In this study, we transferred cultured rabbit cumulus cells (RCC) and fetal fibroblasts (RFF) from genetically marked rabbits (Ali/Bas) into metaphase II (MII) oocytes and analyzed the levels of histone H3K9 acetylation in donor cells and cloned embryos. We also assessed the correlation between the histone acetylation status of donor cells and cloned embryos and their developmental potential. To test whether alteration of the histone acetylation status affects development of cloned embryos, we treated donor cells with sodium butyrate (NaBu), a histone deacetylase inhibitor. Further, we tried to improve cloning efficiency by chimeric complementation of cloned embryos with one or two blastomeres from in vitro-fertilized or parthenogenetic embryos. Histone acetylation in donor cells and cloned embryos was detected by anti-acH3K9 antibody using Western immunoblot analysis or immunochemistry, respectively. Data were analyzed by chi-square (developmental rates) or Student-Newman-Keuls (histone acetylation) test. The levels of acetylated histone H3K9 were higher in RCCs than in RFFs (P &lt; 0.05). Although the type of donor cells did not affect development to blastocyst, after transfer into recipients, RCC-cloned embryos induced a higher initial pregnancy rate as compared to RFF-cloned embryos (40% vs. 20%; P &lt; 0.05). However, almost all pregnancies with either type of cloned embryos were lost by the middle of gestation and only one fully developed; a live RCC-derived rabbit was obtained. Treatment of RFFs with NaBu significantly (P &lt; 0.05) increased the level of histone H3K9/14 acetylation and the proportion of nuclear transfer embryos developing to blastocyst (49% vs. 33% with non-treated RFF; P &lt; 0.05). The distribution of signals for acH3K9 in either group of cloned embryos did not resemble that in in vivo-fertilized embryos, suggesting that reprogramming of this epigenetic mark is aberrant in cloned rabbit embryos and cannot be corrected by treatment of donor cells with NaBu. Aggregation of embryos cloned from NaBu-treated RFFs with blastomeres from in vivo-derived embryos improved development to blastocyst, but no cloned offspring were obtained. Two live cloned rabbits were produced from this donor cell type only after aggregation of cloned embryos with a parthenogenetic blastomere. Our study demonstrates that the levels of histone acetylation in donor cells and cloned embryos correlate with their developmental potential and can be a useful epigenetic mark to predict efficiency of SCNT rabbits. This work was supported by the Bayerische Forschungsstiftung and by Therapeutic Human Polyclonals, Inc.

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