Basic features of bovine spermatogonial culture and effects of glial cell line-derived neurotrophic factor

Aponte, Pedro M.; Soda, Takeshi; Kant, H. J. G. van de; Rooij, Dirk G. de

doi:10.1016/j.theriogenology.2005.10.020

Cited by 81 publications

(79 citation statements)

References 38 publications

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“…Using qPCR, we further confirmed the expression of mRNA encoding other SSC specific markers (viz., β1 integrin and Oct-4) required for colonization and self renewal of SSCs [54,55]. A relatively higher abundance of vimentin mRNA observed in the study might be due to the fact that most SSC cultures contain a mixture of testicular cells constituting as less as 1.33 % SSCs and other testicular cells mainly comprising of Sertoli cells [26].…”

Section: Discussionmentioning

confidence: 66%

“…Morphological and immnocytochemistry analysis revealed that the SSC colonies are intermingled with the somatic cells as some colonies also expressed vimentin. Earlier studies also reported densely packed somatic cell areas in SSC colonies [11,26]. Vimentin, an intermediate filament of Sertoli cell cytoskeleton, plays an important role in maintenance of spermatogenesis in the testes [53].…”

Section: Discussionmentioning

confidence: 81%

“…Bovine SSCs were successfully cultured on a monolayer of STO (SIM 6-thioguanine resistant ouabain resistant) feeder cells [24], bovine embryonic fibroblast cells (BEF) [25] and Sertoli cells [26]. Bovine SSCs were also cultured for short term on laminin coated plates in vitro [11].…”

Section: Introductionmentioning

confidence: 99%

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In vitro culture and characterization of spermatogonial stem cells on Sertoli cell feeder layer in goat (Capra hircus)

Pramod

Mitra

2014

J Assist Reprod Genet

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Purpose To develop an efficient protocol for isolation, purification and long-term culture of spermatogonial stem cell (SSC) in goat. Methods The isolation of SSC was performed by testicular disaggregation by enzymatic digestion using collagenase IV, trypsin and DNase I. Further SSCs were enriched using Percoll density gradient centrifugation. The purity of SSCs was assessed by immunocytochemistry (ICC) using α6 integrin. The SSCs were co-cultured on Sertoli cell feeder layer. The SSC colonies were characterized by studying the expression of SSC specific markers (viz., α6 integrin and PLZF) using ICC. The abundance of mRNAs encoding the markers of SSC (viz., β1 integrin and Oct-4) and Sertoli cells (viz., vimentin) was also assayed using quantitative real-time PCR (qPCR). Results The viability of isolated testicular cells was>90 % and the Percoll density gradient method resulted in 3.65 folds enrichment with a purity of 82.5 %. Co-culturing of SSCs with Sertoli cell feeder layer allowed the maintenance of stable SSC colonies even after one and half months of culture. The results of ICC analysis showed the expression of α6 integrin and PLZF in almost all the SSC colonies. qPCR analysis revealed a differential expression of mRNAs encoding β1 integrin, Oct-4 and vimentin markers. Conclusion Results of this study demonstrate a simple enzymatic digestion and Percoll density gradient method for isolation and enrichment of SSCs, and suitability of Sertoli cell feeder layer for long term in vitro culture of SSC in goats.Results also suggest the possible application of non-caprine antibodies against SSC specific markers for the identification and subsequent assessment of SSCs in goats.

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Section: Discussionmentioning

confidence: 66%

Section: Discussionmentioning

confidence: 81%

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In vitro culture and characterization of spermatogonial stem cells on Sertoli cell feeder layer in goat (Capra hircus)

Pramod

Mitra

2014

J Assist Reprod Genet

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“…Binding of VEGF to its cognate receptors initiates a variety of signaling cascades similar to those stimulated by GDNF and KIT in the postnatal testis, two factors of known importance for regulating germ cell proliferation, VEGFA-164 regulates germ cell survival survival, and differentiation (Meng et al 2000, Ohta et al 2000, Rossi et al 2000, Dolci et al 2001, Aponte et al 2006, Naughton et al 2006, Payne & Braun 2007. Rodent studies have shown that male fetal germ cells express VEGF protein until shortly after birth (Bott et al 2006), and Sertoli cells continue to produce VEGF throughout adult life (Marti & Risau 1998, Nalbandian et al 2003.…”

Section: Discussionmentioning

confidence: 99%

Vascular endothelial growth factor regulates germ cell survival during establishment of spermatogenesis in the bovine testis

Caires

Avila²,

McLean

2009

Reproduction

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Vascular endothelial growth factor-A (VEGFA) is a hypoxia-inducible peptide essential for angiogenesis and targets nonvascular cells in a variety of tissues and cell types. The objective of the current study was to determine the function of VEGF during testis development in bulls. We used an explant tissue culture and treatment approach to test the hypothesis that VEGFA-164 could regulate the biological activity of bovine germ cells. We demonstrate that VEGFA, KDR, and FLT1 proteins are expressed in germ and somatic cells in the bovine testis. Treatment of bovine testis tissue with VEGFA in vitro resulted in significantly more germ cells following 5 days of culture when compared with controls. Quantitative real-time RT-PCR analysis determined that VEGF treatment stimulated an intracellular response that prevents germ cell death in bovine testis tissue explants, as indicated by increased expression of BCL2 relative to BAX and decreased expression of BNIP3 at 3, 6, and 24 h during culture. Blocking VEGF activity in vitro using antisera against KDR and VEGF significantly reduced the number of germ cells in VEGF-treated testis tissue to control levels at 120 h. Testis grafting provided in vivo evidence that bovine testis tissue treated with VEGFA for 5 days in culture contained significantly more differentiating germ cells compared with controls. These findings support the conclusion that VEGF supports germ cell survival and sperm production in bulls.

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“…Thus, increasing proliferation and viability rate of SSCs in vitro facilitates great successes in the field of genetic improvement of cattle herds. Type A spermatogonia can be obtained in large numbers in prepubertal bulls (4,5). Isolation of type A spermatogonia is performable by 2-enzyme digestion of calf testes, which gives 65%-87% pure cell populations (6).…”

Section: Introductionmentioning

confidence: 99%

Study of insulin-like growth factor 1 effects on bovine type A spermatogonia proliferation and viability

panahi¹,

Tajik²,

Movahedin³

et al. 2014

Turk J Vet Anim Sci

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In this survey, 5 male Holstein Friesian calves of 3 months old were used. The research was conducted in accordance with National Research Council guidelines. Calves were sedated with xylazine (0.1 mg/kg of body weight, i.m.) and received a 3-mL injection of 2% lidocaine 20 min before surgery. Respiration rate, heart rate, and rectal temperature were monitored before and after recovery from analgesia. Calves were given flunixin meglumine (2.2 mg/kg of body weight, i.m.) for 3 days after surgery.

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Basic features of bovine spermatogonial culture and effects of glial cell line-derived neurotrophic factor

Cited by 81 publications

References 38 publications

In vitro culture and characterization of spermatogonial stem cells on Sertoli cell feeder layer in goat (Capra hircus)

In vitro culture and characterization of spermatogonial stem cells on Sertoli cell feeder layer in goat (Capra hircus)

Vascular endothelial growth factor regulates germ cell survival during establishment of spermatogenesis in the bovine testis

Study of insulin-like growth factor 1 effects on bovine type A spermatogonia proliferation and viability

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