Configuration and distribution of bovine spermatogonia

Wrobel, Karl‐Heinz; Bickel, Daniela; Kujat, Richard; Schimmel, Margit

doi:10.1007/bf00318484

Cited by 37 publications

(32 citation statements)

References 18 publications

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“…In the current study nearly all large type A spermatogonia in the immature goat testes were c-kit-positive, while the subpopulations of small (basal) spermatogonia were c-kitnegative. This finding confirmed the classification of Wrobel et al (1995) in which the smaller spermatogonia were less differentiated than larger ones [24]. In this agreement in mice there is little expression of the c-kit mRNA and protein in undifferentiated type A spermatogonia and its expression is clearly enhanced upon differentiation of these cells into type A1 spermatogonia.…”

Section: Discussionsupporting

confidence: 90%

Isolation, identification, and culture of goat spermatogonial stem cells using c-kit and PGP9.5 markers

Heidari

Rahmati-Ahmadabadi

Akhondi

et al. 2012

J Assist Reprod Genet

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Introduction Presently the techniques for making transgenic animals are cumbersome, required costly instruments and trained man-power. The ability of spermatogonial stem cells (SSCs) to integrate foreign genes has provided the opportunity for developing alternate methods for generation of transgenic animals. One of the big challenges in this field is development of the methods to identify and purify donor SSCs by antibody mediated cell sorting. Purpose The present study was aimed to identify goat subpopulations of SSCs using polyclonal antibodies against PGP9.5 and c-kit molecular markers as well as the growth characteristics of SSCs during short term culture. Methods One month old goats' testicular samples were subjected for immunohistochemical and immunocytochemical evaluations. The enzymatically isolated SSCs were cultured in DMEM plus FCS supplemented with (treatment) or without (control) growth factors (GDNF, LIF, FGF, and EGF) for 2 weeks. At the end of culture the morphological characteristics of SSCs colonies and immunocytochemical staining were evaluated. Results The number and size of colonies in treatment groups were significantly (P<0.01) higher than corresponding values in controls. The presence of PGP 9.5 and c-kit antigens was confirmed in immunocytochemical evaluation. In immunocytochemical evaluation, the proportion of c-kit and PGP9.5 positive cells were significantly (P<0.001) higher in control and treatment groups, respectively. Conclusions The presence of PGP9.5 and c-kit antigens was confirmed in goat SSCs. Moreover, culture medium supplementation with growth factors could effectively retain the undifferentiation status of SSCs, reflected as a higher population of PGP9.5 positive cells, after short term culture.

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Section: Discussionsupporting

confidence: 90%

Isolation, identification, and culture of goat spermatogonial stem cells using c-kit and PGP9.5 markers

Heidari

Rahmati-Ahmadabadi

Akhondi

et al. 2012

J Assist Reprod Genet

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“…Spermatogonia were identified using protein gene product 9.5 antigen (PGP 9.5, Chemicon Australia Pty Ltd, Melbourne, Australia) in Bouin's fixed testis sections from the donor animals and also in the cell suspensions after overnight culture (Wrobel et al 1995, Rathi et al 2005. Slides were quenched by incubating in methanol-peroxide for 5 min to block the endogenous peroxidase activity, permeabilized with Trisbuffered saline tween-20C0.01% Triton X-100 for 5 min and blocked with 0.5% BSA in TBS for 30 min.…”

Section: Assessment Of Spermatogonia Populationmentioning

confidence: 99%

Successful transplantation of bovine testicular cells to heterologous recipients

Herrid¹,

Vignarajan²,

Davey³

et al. 2006

Reproduction

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While heterologous germ cell transplantation was successful in pigs and goats, autologous transplantation alone has been reported to result in donor-derived spermatogenesis in cattle. The objective of this study was to investigate whether the transplantation of heterologous germ cells could result in colonization of recipient testes in cattle of different breeds. Testicular cells were isolated from 8 Bos taurus donor bull calves and then transferred into 15 Bos indicus-cross bull calves. All animals were pre-pubertal, donors were aged 5-7 months and recipients 5-11 months, and scrotal circumferences ranged from 15 to 22 cm. Single cell suspensions of donor testicular cells, prepared by enzymatic digestion, were labelled with fluorescent dyes PKH26 or CFDA-SE, before transfer into the rete testis of recipients under ultrasonographic guidance. To assess the longevity of colonization by donor cells, recipients were castrated 2-30 weeks after cell transfer. Donor cells were observed in 15/25 (60%) of the testes that received PKH26-labelled cells, whereas no CFDA-SE-positive cell was identified in any recipients. The maturity of the donors or recipients (measured by scrotal circumference) did not affect colonization potential. In freshly isolated tubules, clumps of PKH26-positive cells were observed, which indicated either cell division or extensive local colonization of specific areas of the tubules. In frozen sections, PKH26-positive cells were identified on the seminiferous tubule basement membrane, which indicated that these cells had successfully migrated from the tubule lumen and were likely to be spermatogonia. We conclude that PKH26 was more suitable for labelling donor testis cells and donor cells can be identified up to 6 months following transfer. These results indicate that allogeneic transplantation of testicular cells can occur between Bos taurus and Bos indicus cattle. Further studies will investigate functionality of transferred testicular cells.

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“…Immunohistochemistry of protein gene product 9.5 (PGP 9.5) PGP 9.5 immunohistochemistry was used to identify gonocytes and spermatogonia (Wrobel et al 1995) in the tissue samples. The protocol was modified from Wrobel et al (1995).…”

Section: Recovery and Analysis Of Xenograftsmentioning

confidence: 99%

“…The protocol was modified from Wrobel et al (1995). Briefly, slides were de-paraffinized and tissue samples were treated with 3% H 2 O 2 in distilled water for 10 min to block the endogenous peroxidase activity followed by two washes in PBS for 5 min each.…”

Section: Recovery and Analysis Of Xenograftsmentioning

confidence: 99%

Germ cell fate and seminiferous tubule development in bovine testis xenografts

Rathi¹,

Honaramooz²,

Zeng³

et al. 2005

Reproduction

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Spermatogenesis can occur in testis tissue from immature bulls ectopically grafted into mouse hosts; however, efficiency of sperm production is lower than in other donor species. To elucidate a possible mechanism for the impaired spermatogenesis in bovine testis xenografts, germ cell fate and xenograft development were investigated at different time points and compared with testis tissue from age-matched calves as controls. Histologically, an initial decrease in germ cell number was noticed in xenografts recovered up to 2 months post-grafting without an increase in germ cell apoptosis. From 2 months onward, the number of germ cells increased. In contrast, a continuous increase in germ cell number was seen in control tissue. Pachytene spermatocytes were observed in some grafts before 4 months, whereas in the control tissue they were not present until 5 months of age. Beyond 4 months post-grafting spermatogenesis appeared to be arrested at the pachytene spermatocyte stage in most grafts. Elongated spermatids were observed between 6 and 8 months post-grafting, similar to the controls, albeit in much lower numbers. Lumen formation started earlier in grafts compared with controls and by 6 months post-grafting tubules with extensively dilated lumen were observed. A donor effect on efficiency of spermatogenesis was also observed. These results indicate that the low efficiency of sperm production in bovine xenografts is due to an initial deficit of germ cells and impaired meiotic and post-meiotic differentiation. The characterization of spermatogenic efficiency will provide the basis to understand the control of spermatogenesis in testis grafts.

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Configuration and distribution of bovine spermatogonia

Cited by 37 publications

References 18 publications

Isolation, identification, and culture of goat spermatogonial stem cells using c-kit and PGP9.5 markers

Isolation, identification, and culture of goat spermatogonial stem cells using c-kit and PGP9.5 markers

Successful transplantation of bovine testicular cells to heterologous recipients

Germ cell fate and seminiferous tubule development in bovine testis xenografts

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