Microsatellite detection of donor-derived sperm DNA following germ cell transplantation in cattle

2009 ◽  
Vol 21 (3) ◽  
pp. 462 ◽  
Author(s):  
Sally Stockwell ◽  
Muren Herrid ◽  
Rhonda Davey ◽  
Alan Brownlee ◽  
Keryn Hutton ◽  
...  
Keyword(s):  
Germ Cell ◽  
Microsatellite Markers ◽  
Cell Transplantation ◽  
Breeding Systems ◽  
Testicular Germ Cell ◽  
Livestock Breeding ◽  
Germ Cell Transplantation ◽  
Sperm Dna ◽  
Heterologous Transplantation ◽  
Potential Applications

Although autologous and heterologous transplantation has resulted in colonisation of recipient testes in cattle, the ability of the transplanted spermatogonial stem cells to complete spermatogenesis has not yet been determined. The objective of the present study was to identify and validate microsatellite markers that can distinguish the genotype of different individuals and therefore can be used to detect the presence of donor DNA in recipient semen samples. In a previous study by this group, successful colonisation of recipient testes by heterologous transfer using a fluorescent dye was shown. In the present work, some of the same recipient animals were investigated further to monitor donor-derived sperm production. The bovine microsatellite detection method was developed specifically to test the ejaculates of the recipients and can also be used to pre-match individuals before germ cell transplantation. Semen was collected from the recipients 52–98 weeks after transfer and the presence of donor DNA in the samples was determined using microsatellite markers. In one of the recipients, all collected semen samples were shown to be positive for donor-derived cells; however, the percentage of donor spermatozoa in the recipient ejaculate declined with time. The donor DNA was also detected in both single cell suspensions and testis tissue from this recipient. These results demonstrate for the first time that testicular germ cell transplantation between different breeds of cattle is feasible and the recipients thereof are able to produce spermatozoa of donor origin. This technology has potential applications in livestock breeding systems and may provide an alternative to artificial insemination.

269. Optimal testicular size of donor and recipient for testicular germ cell transplantation in the bovine

2005 ◽  
Vol 17 (9) ◽  
pp. 110
Author(s):  
M. Herrid ◽  
R. Davey ◽  
S. Vignarajan ◽  
K. Hutton ◽  
J. Hill
Keyword(s):  
Germ Cell ◽  
Cell Transplantation ◽  
Cell Number ◽  
Rete Testis ◽  
Colonization Rate ◽  
Cell Transplant ◽  
Testicular Germ Cell ◽  
Scrotal Circumference ◽  
Germ Cell Transplantation ◽  
Donor Cells

The maturity status of donor and recipient testis appears to be important in the efficiency of testicular germ cell transplantation. When neonatal mice were used as recipients, they show 9.4 times greater colonization and 4 times larger colony size than in the adult.1 The objective of this study in cattle was to investigate the effect of testicular maturity of donor and recipient calves on success of the testis cell transplant procedure. Testicular maturity was measured indirectly by scrotal circumference and donor testicular cells were enzymatically isolated from 8 Angus calves aged 5–7 months (scrotal circumference, SC of 18–22cm) and injected into both testes of 12 recipient calves aged 4–6 months (SC 15–21cm. Donor cells were labeled with the red fluorescent dye PKH26 then transferred into the rete testis under ultrasonographic guidance. Castration of recipients was performed 2–6 months following injection and then frozen sections were used to localize the PKH26 positive donor cells. Five sections from different 5 areas in each testis were prepared and 100 tubules were counted. In 15 of the 24 (63%) testes, PKH positive donor cells were identified. There was no correlation between colonization rate and maturity of donor animal testis for the range of testis sizes studied. Testis cells from donors of SC 18-20 cm or of SC 21-22 did not result in different number of recipient testis with positive cells (7/10 (70%) v. 8/14 (57%)) or the number of positive cells per testis (1.92 ± 0.67% v. 2.5 ± 1.01%). Recipient maturity (SC of 15–18 cm v. SC of 19–21 cm) had no effect on the colonization rate (7/11 (64%) v. 8/13 (62%)); however, there were significantly more positive cells per testis in less mature (SC of 15–18) recipients (3.18±1.21% v. 1.52 ± 0.64% P < 0.05)). In summary we have demonstrated successful testicular germ cell transplantation between calves and while donor testis cell age appeared to have little effect on the efficiency of colonization, less mature testis provided more suitable conditions for colonization. (1)Shinohara T, Orwig KE, Avarbock MR and Brinster RL. (2001) Remodeling of the postnatal mouse testis is accompanied by dramatic changes in stem cell number and niche accessibility. PNAS 98(11), 6186–6191.

scholarly journals Irradiation Enhances the Efficiency of Testicular Germ Cell Transplantation in Sheep

2009 ◽  
Vol 81 (5) ◽  
pp. 898-905 ◽  
Author(s):  
Muren Herrid ◽  
Jeanette Olejnik ◽  
Michael Jackson ◽  
Natalka Suchowerska ◽  
Sally Stockwell ◽  
...  
Keyword(s):  
Germ Cell ◽  
Cell Transplantation ◽  
Testicular Germ Cell ◽  
Germ Cell Transplantation

Irradiation Enhanced Efficiency of Testicular Germ Cell Transplantation in Sheep.

2008 ◽  
Vol 78 (Suppl_1) ◽  
pp. 121-122
Author(s):  
Muren Herrid ◽  
Michael Jackson ◽  
Natalka Suchowerska ◽  
Sally Stockwell ◽  
Keryn Hutton ◽  
...  
Keyword(s):  
Germ Cell ◽  
Cell Transplantation ◽  
Testicular Germ Cell ◽  
Germ Cell Transplantation

Donor sperm production in heterologous recipients by testis germ cell transplantation in the dromedary camel

2019 ◽  
Vol 31 (3) ◽  
pp. 538 ◽  
Author(s):  
Muren Herrid ◽  
Peter Nagy ◽  
Jutka Juhasz ◽  
Jane M. Morrell ◽  
M. Billah ◽  
...  
Keyword(s):  
Germ Cell ◽  
Cell Transplantation ◽  
Gradient Centrifugation ◽  
Donor Cell ◽  
Fold Increase ◽  
Rete Testis ◽  
Dromedary Camel ◽  
Testicular Germ Cell ◽  
Donor Sperm ◽  
Germ Cell Transplantation

The object of this study was to investigate if testis germ cell transplantation (TGCT) into a heterologous recipient would result in donor-origin spermatogenesis in the dromedary camel. First, we investigated a workable protocol for TGCT in camels, including donor cell isolation, enrichment by density gradient centrifugation (Percoll and Bovicoll), rete testis injection and microsatellite detection of donor and recipient genotypes. Second, the effects of three doses of Dolichos biflorus agglutinin (DBA), a glycoprotein that specifically binds to gonocytes or Type A spermatogonia, on testis germ cell depletion were investigated by direct injection into the rete testis of a male camel. Seven recipients were prepared with DBA treatment, two males were castrated at 4 weeks for depletion assessment and the remaining five received donor cells 4–6 weeks after treatment. On average, ~17 million cells were isolated per gram of testis tissue, with 19.5±1.9% DBA-positive (DBA+) cells. Percoll centrifugation yielded a 1.5-fold increase in DBA+ cells while Bovicoll centrifugation produced a 2.5-fold increase from the input cells of 18.6±2.1% DBA+ cells. Semen was collected from the recipients 13–20 weeks after transfer and the presence of donor DNA in the samples was determined using microsatellite markers. In two of the five recipients, all semen samples were shown to be positive for donor-derived cells. These results demonstrate for the first time that: (1) heterologous testicular germ cell transplantation in camels is feasible and the recipients are able to produce spermatozoa of donor origin and (2) DBA can be used effectively to deplete endogenous stem cells.

scholarly journals Induced sterility in fish and its potential and challenges for aquaculture and germ cell transplantation technology: a review

Biologia ◽  
2016 ◽  
Vol 71 (8) ◽  
Author(s):  
Amin Golpour ◽  
Mohammad Abdul Momin Siddique ◽  
Diógenes Henrique Siqueira-Silva ◽  
Martin Pšenička
Keyword(s):  
Germ Cell ◽  
Cell Transplantation ◽  
Large Scale ◽  
Hormonal Treatment ◽  
Important Species ◽  
Genetic Ablation ◽  
Germ Cell Transplantation ◽  
Commercially Important ◽  
Alternative Approaches ◽  
Hormone Signalling

AbstractInterest in reproductively sterile fish in aquaculture has prompted research into their production. Several methods are available for inducing sterility and optimizing its application in the global fishery industry. Sterilization can potentially be accomplished through irradiation, surgery, or chemical and hormonal treatment. Alternative approaches include triploidization, hybridization, and generation of new lines via advanced biotechnological techniques. Triploids of many commercially important species have been studied extensively and have been produced on a large scale for many years. Novel approaches, including disruption of gonadotropin releasing hormone signalling and genetic ablation of germ cells, have been developed that are effective in producing infertile fish but have the disadvantage of not being 100% reliable or are impractical for large-scale aquaculture. We review currently used technologies and recent advances in induction of sterility in fish, especially those intended for use in germ cell transplantation. Knowledge of the implications of these approaches remains incomplete, imposing considerable limitations.

scholarly journals Restoration of Fertility by Germ Cell Transplantation Requires Effective Recipient Preparation1

2003 ◽  
Vol 69 (2) ◽  
pp. 412-420 ◽  
Author(s):  
Clayton J. Brinster ◽  
Buom-Yong Ryu ◽  
Mary R. Avarbock ◽  
Levent Karagenc ◽  
Ralph L. Brinster ◽  
...  
Keyword(s):  
Germ Cell ◽  
Cell Transplantation ◽  
Germ Cell Transplantation ◽  
Restoration Of Fertility

scholarly journals Male germ cell transplantation in livestock

2006 ◽  
Vol 18 (2) ◽  
pp. 13 ◽  
Author(s):  
J. R. Hill ◽  
I. Dobrinski
Keyword(s):  
Stem Cells ◽  
Germ Cell ◽  
Cell Transplantation ◽  
Large Scale ◽  
Developmental State ◽  
Male Germ Cell ◽  
Seminiferous Tubules ◽  
Cell Transfer ◽  
Donor Sperm ◽  
Germ Cell Transplantation

Male germ cell transplantation is a powerful approach to study the control of spermatogenesis with the ultimate goal to enhance or suppress male fertility. In livestock animals, applications can be expanded to provide an alternative method of transgenesis and an alternative means of artificial insemination (AI). The transplantation technique uses testis stem cells, harvested from the donor animal. These donor stem cells are injected into seminiferous tubules, migrate from the lumen to relocate to the basement membrane and, amazingly, they can retain the capability to produce donor sperm in their new host. Adaptation of the mouse technique for livestock is progressing, with gradual gains in efficiency. Germ cell transfer in goats has produced offspring, but not yet in cattle and pigs. In goats and pigs, the applications of germ cell transplantation are mainly in facilitating transgenic animal production. In cattle, successful male germ cell transfer could create an alternative to AI in areas where it is impractical. Large-scale culture of testis stem cells would enhance the use of elite bulls by providing a renewable source of stem cells for transfer. Although still in a developmental state, germ cell transplantation is an emerging technology with the potential to create new opportunities in livestock production.

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