Isolation and transplantation of spermatogonia in sheep

Rodriguez‐Sosa, Jose R.; Dobson, Howard; Hahnel, Ann

doi:10.1016/j.theriogenology.2006.03.039

Cited by 93 publications

(85 citation statements)

References 35 publications

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“…In Rodriguez-Sosa et al (2006), we found that the spermatogonial fraction (Fraction A) contains on average 65% UCHL1 (PGP 9.5) positive spermatogonia (65.3G3.2%), and the 'somatic' fraction (Fraction B) contains w80% vimentin and smooth muscle actin positive, peritubular myoid cells (80.8G3.2%). Cells from each fraction were frozen according to Izadyar et al (2002).…”

Section: Spermatogonial Transplantationmentioning

confidence: 99%

“…Spermatogonial and somatic cell fractions were purified from two 45 cm 3 testes (see above) by a two step enzymatic procedure and Percoll gradients as previously described (Rodriguez-Sosa et al 2006). In Rodriguez-Sosa et al (2006), we found that the spermatogonial fraction (Fraction A) contains on average 65% UCHL1 (PGP 9.5) positive spermatogonia (65.3G3.2%), and the 'somatic' fraction (Fraction B) contains w80% vimentin and smooth muscle actin positive, peritubular myoid cells (80.8G3.2%).…”

Section: Spermatogonial Transplantationmentioning

confidence: 99%

“…Cells from each fraction were frozen according to Izadyar et al (2002). On the day of xenografting, these cells were thawed at 37 8C for 1-2 min and labeled with the cell tracking dye CFDA (Molecular Probes, Eugene, OR, USA) as previously described (Rodriguez-Sosa et al 2006). A total of 150!10 6 cells of each type (spermatogonial or somatic) were suspended in 3 ml of 0.04% trypan blue in sterile PBS.…”

Section: Spermatogonial Transplantationmentioning

confidence: 99%

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Development of strips of ovine testes after xenografting under the skin of mice and co-transplantation of exogenous spermatogonia with grafts

Rodriguez‐Sosa

Foster²,

Hahnel³

2010

Reproduction

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Xenografting of testicular tissue is an attractive new strategy for studying postnatal development of spermatogenesis and to preserve male genetics in large mammals. Typically, small cubes of immature testis (1 mm 3 ) are grafted under the dorsal skin of immune-deficient mice. We attempted to increase the total number of seminiferous tubules in each xenograft with spermatogenesis by grafting flat strips of testis (w9!5!1 mm) from ram lambs in immune-deficient mice. The percentage of grafts that survived and percentage of seminiferous tubules that developed spermatogenesis were the same as those reported after xenografting small cubes of lamb testis. Partially purified sheep spermatogonia were labeled with the fluorescent dye carboxy fluorescein diacetate succinyl diester and transplanted into the seminiferous tubules of one of the donor testis just before engraftment. The temporary label in the donor cells was detected for 4 weeks after xenografting, suggesting that co-engraftment of spermatogonia with testicular tissue may be a way to rapidly determine the effect of a specific gene on spermatogenesis. Finally, Sertoli cell lesions in xenografts of lamb testes were quantified, and their number and severity were found to increase, especially after grafts had been in place for 4 weeks. Although this coincided with the development of spermatogenesis, the extent of germ cell differentiation negatively correlated with severity of the lesions.

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Section: Spermatogonial Transplantationmentioning

confidence: 99%

Section: Spermatogonial Transplantationmentioning

confidence: 99%

Section: Spermatogonial Transplantationmentioning

confidence: 99%

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Development of strips of ovine testes after xenografting under the skin of mice and co-transplantation of exogenous spermatogonia with grafts

Rodriguez‐Sosa

Foster²,

Hahnel³

2010

Reproduction

Self Cite

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“…One such a marker is c-kit, the receptor for stem cell factor (SCF), which is expressed by some Aal, A1-A4, In, and B spermatogonia [12]. The presence of c-kit receptor in SSCs, however, has been a matter of debate for many years [13,14].…”

Section: Introductionmentioning

confidence: 99%

“…Antibody against this protein has been used for immunohistochemical detection of mouse [15], bovine [16], ram [14], human [17] and porcine [18] spermatogonia. Respecting to the lack of information on goat type A spermatogonial markers and the inadequate information regarding to the isolation, identification, and culture of goat SSCs, and considering the application of transfected SSCs as an efficient tool in production of transgenic animals, the present study was aimed to investigate the identification of prepubertal goat SSCs using polyclonal antibodies against PGP9.5 and c-kit molecular markers.…”

Section: Introductionmentioning

confidence: 99%

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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Recent advances of in vitro culture systems for spermatogonial stem cells in mammals

Sahare

Suyatno

Imai

2018

Reprod Medicine & Biology

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BackgroundSpermatogonial stem cells (SSCs) in the mammalian testis are unipotent stem cells for spermatozoa. They show unique cell characteristics as stem cells and germ cells after being isolated from the testis and cultured in vitro. This review introduces recent progress in the development of culture systems for the establishment of SSC lines in mammalian species, including humans.MethodsBased on the published reports, the isolation and purification of SSCs, identification and characteristics of SSCs, and culture system for mice, humans, and domestic animals have been summarized.ResultsIn mice, cell lines from SSCs are established and can be reprogrammed to show pluripotent stem cell potency that is similar to embryonic stem cells. However, it is difficult to establish cell lines for animals other than mice because of the dearth of understanding about species‐specific requirements for growth factors and mechanisms supporting the self‐renewal of cultured SSCs. Among the factors that are associated with the development of culture systems, the enrichment of SSCs that are isolated from the testis and the combination of growth factors are essential.ConclusionProviding an example of SSC culture in cattle, a rational consideration was made about how it can be possible to establish cell lines from neonatal and immature testes.

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Isolation and transplantation of spermatogonia in sheep

Cited by 93 publications

References 35 publications

Development of strips of ovine testes after xenografting under the skin of mice and co-transplantation of exogenous spermatogonia with grafts

Development of strips of ovine testes after xenografting under the skin of mice and co-transplantation of exogenous spermatogonia with grafts

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

Recent advances of in vitro culture systems for spermatogonial stem cells in mammals

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