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.

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.

251. Production of donor-derived live lambs following testis germ cell transplantation

2008 ◽  
Vol 20 (9) ◽  
pp. 51
Author(s):  
M. Herrid ◽  
M. Jackson ◽  
N. Suchowerska ◽  
S. Stockwell ◽  
K. Hutton ◽  
...  
Keyword(s):  
Germ Cell ◽  
Cell Transplantation ◽  
Single Cells ◽  
Pubertal Stage ◽  
No Donor ◽  
Post Irradiation ◽  
Germ Cell Transplantation ◽  
Donor Cells ◽  
The Right ◽  
Sperm Output

Testes germ cell transplantation in livestock has the potential for amplification of transgenic genotypes and for use as an alternative to artificial insemination. This study investigated a workable protocol for testis germ cell transplantation in sheep between animals of the same breed and different breeds. Testes of two groups of recipients at the stage of pre-pubertal (transition from gonocytes to spermatogonia, n = 2) or peri-pubertal (spermatogenesis initiated, n = 2) were treated with a single dose of 9 grey (Gy) or 15 Gy with a 6MV photon beam irradiation, respectively. In the first experiment, using pre-pubertal irradiated animals, testis germ cell transplantation between the same breed was performed at 16 weeks post irradiation. The left testes of recipient rams were injected with donor cells labelled with fluorescent dye PHK26, while the right testes were given unlabelled cells. The left testes of recipients were removed by castration after 2 weeks following transplantation to evaluate the location of the transferred cells, while the right testes were kept in place for long-term assessment of sperm output. In cryosections of the left testes, PKH26 positive cells were found both on the basement membrane as single cells or in the interstitial area. In the second experiment, animals irradiated at the peri-pubertal stage, received donor cells at 5 weeks post irradiation and animals were kept intact for semen production. For a period of two years after transplantation, semen samples were collected routinely from two groups of rams and analysed using microsatellite markers. Two recipients (50%) demonstrated the presence of donor DNA in their ejaculates. In order to investigate the fertility of the donor-origin sperm in recipient ejaculates, 99 ewes were artificially inseminated with semen from two positive rams. Four lambs (8%) have been identified as being sired by donor-derived sperm produced in the recipient ram that received a Merino to Merino transplantation, while no donor-derived offspring was obtained from the recipient with Border Leicester to Merino transplantation. This study represents the first report of the production of live progeny following testis germ cell transplantation in sheep.

Male germ cell transplantation: promise and problems

2001 ◽  
Vol 13 (8) ◽  
pp. 609 ◽  
Author(s):  
Fang-Xu Jiang
Keyword(s):  
Germ Cell ◽  
Cell Transplantation ◽  
Germ Cells ◽  
Cell Biology ◽  
Direct Injection ◽  
Primordial Germ Cells ◽  
Rete Testis ◽  
Male Germ Cell ◽  
Seminiferous Tubules ◽  
Germ Cell Transplantation

Male germ cell transplantation is a novel technique in which donor male stem germ cells are surgically transferred to the seminiferous tubules of a recipient testis by direct injection or via the rete testis or efferent duct. All germ cells that are destined to become stem spermatogonia are defined as male stem germ cells, including primordial germ cells from the gonadal ridges, and gonocytes and stem spermatogonia from the testis, all of which are transplantable and capable of undergoing normal spermatogenesis. Xenotransplantation of male germ cells from one species into the testis of another species, including human testicular cells in the mouse, has so far proved to be unsuccessful. However, the immunodeficient mouse testis can support rat spermatogenesis and produce apparently normal rat spermatozoa. The underlying mechanisms remain elusive. The present mini-review will focus on the importance of stem spermatogonial transplantation for testicular stem cell biology and discuss the likelihood of immune rejection after transplantation, which may limit the success of all male germ cell transplantation.

scholarly journals MORPHOLOGICAL CHARACTERISTIC OF SPERMATOGONIA AND TESTES DISSOCIATION : A Preliminary Study for the Germ Cell Transplantation in Giant Gouramy (Osphronemus gouramy)

2010 ◽  
Vol 5 (2) ◽  
pp. 163
Author(s):  
Irma Andriani ◽  
Ita Djuwita ◽  
Komar Sumantadinata ◽  
Muhammad Zairin Junior ◽  
Harton Arfah ◽  
...  
Keyword(s):  
Germ Cell ◽  
Cell Transplantation ◽  
Incubation Time ◽  
Trypan Blue Exclusion ◽  
Endangered Fish ◽  
Germ Cell Transplantation ◽  
Donor Cells ◽  
Viable Population ◽  
Preliminary Study ◽  
Optimal Incubation

The recent study were attempting to develop spermatogonial germ cell transplantation as a tool to preserve and propagate male germ-plasm from endangered fish species, as well as to produce surrogate broodstock of commercially valuable fish. Spermatogonia identification and testes dissociation were the first necessary steps to obtain highly amount and viable population of spermatogonia as donor cells for transplantation. Using giant gouramy testes as a model, spermatogonia was histological characterized and two methods of testes dissociations were compared (i.e. medium A contained 0.5% trypsin in PBS and medium B contained 0.5% trypsin and DNase 10 IU/μL in PBS complemented with CaCl2, Hepes and FCS). Optimal incubation times (1, 2, 3, 4 and 5 hours) in dissociation medium were also determined. Freshly isolated testes of immature giant gouramy were minced in dissociation medium and then incubated to get monodisperce cell suspension. Parameters observed were number and viability of spermatogonia (ø > 10 μm). The viability was analyzed using trypan blue exclusion dye. The results showed that the average number of spermatogonia observed in medium B was higher than in medium A (P<0.05), meanwhile the viability of spermatogonia between medium A and B were not significantly different (P>0.05). The viability of spermatogonia decreased by the increasing duration time of dissociation. The viability of spermatogonia started to decrease significantly in 2 hours incubation time in medium A and 4 hours incubation time in medium B (P<0.05). In conclusion, application of dissociation medium B yielded higher number of viable spermatogonia than dissociation medium A.

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.

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

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.

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