Spermatogonial Stem Cell Transplantation in Large Animals

Zhao, Xin; Wan, Weican; Zhang, Xianyu; Wu, Zhenfang; Yang, Huaqiang

doi:10.3390/ani11040918

Cited by 15 publications

(13 citation statements)

References 116 publications

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“…Similarly, in most mammals, including goats, ST exists as a highly coiled structure that is closely packed in testicular lobules [Sarma and Devi, 2017]. Therefore, transplantation of GCs through precise injection into the lumen of ST may require multiple incisions in the tunica albuginea, which increases the complexity of the manipulation and injury to the testis of the recipient [Gul et al, 2020;Zhao et al, 2021]. Furthermore, if not controlled accurately, higher intra-tubular pressure during direct injection into the ST may result in rupture of the tubular membrane and leakage into the interstitium [Gul et al, 2020].…”

Section: Discussionmentioning

confidence: 99%

“…In contrast to these, has a distinct anatomical location of MT in goats that is characterized by a labyrinth of intercommunicating channels of testicular lobules and occupies about two-thirds of the deep central axis of the testis [Goyal and Williams, 1987], making it easily distinct from adjacent tissues and offer more useful access to the ST [Honaramooz et al, 2002]. Moreover, injection into MT allows an infusion of a relatively large volume of solution or cell suspension which can be injected at multiple locations to ensure a higher transfer efficiency [Zhao et al, 2021]. However, consider- ing the location of MT, the use of imaging techniques such as USG are required for a precise injection into the MT [Zhao et al, 2021].…”

Section: Discussionmentioning

confidence: 99%

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Successful in vivo Transplantation of Cultured and Enriched Testicular Germ Cells of Pre-Pubertal Bucks to Busulfan-Treated Homologous Recipients

Singh¹,

Kharche²,

Soni³

et al. 2022

Cells Tissues Organs

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The objective of the present study was to establish a workable approach for the production of germ cell (GC)-depleted recipient goat model using intra-testicular busulfan treatment and transplantation of cultured and enriched caprine-male GC (cmGCs) into the homologous recipients under ultrasonography (USG) guidance. The evaluation of post-transplantation colonization of donor cmGCs and restoration of the normal architecture of seminiferous tubules (ST) was performed. For this, the cmGCs of pre-pubertal male goats were isolated and enriched by differential platting for culture until the third passage. Thereafter, cells were harvested and further enriched by magnetic-activated cell sorting using rabbit-anti-CD90 antibody. After confirmation of metabolic viability (MTT-assay) and cluster-forming ability (crystal violet staining) of CD90<sup>+</sup> cmGCs, the cells were labeled with a lipophilic red-fluorescent dye (PKH26) before transplanted into the recipient male goats by injection directly into the mediastinum testes under USG guidance. The colonization and repopulation of transplanted CD90<sup>+</sup> cmGCs into the recipient ST was observed up to 8 weeks post-transplantation. The PKH26-labeled donor cell-derived colonies were identified in enzymatically digested ST and cryosections of recipient testes. Moreover, histochemical analyses revealed the restoration of the normal architecture of ST of recipient testis after GC transplantation. Therefore, the results suggest that the reproductive competence of infertile animals can be restored through mGC therapy and thus the methodology presented herein could be useful to obtain donor mGCs-derived functional male gametes in the recipient animal testis.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Successful in vivo Transplantation of Cultured and Enriched Testicular Germ Cells of Pre-Pubertal Bucks to Busulfan-Treated Homologous Recipients

Singh¹,

Kharche²,

Soni³

et al. 2022

Cells Tissues Organs

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“…Spermatogenesis is a series of complex processes of sequential cell proliferation and differentiation in male testis, affected by many crucial proteins and signal pathways (15,16). Because SSCs produce a large number of progenitors, they are important in the spermatogenesis process (17).…”

Section: Discussionmentioning

confidence: 99%

The effect of regulating the Wnt signaling pathway on the proliferation and differentiation of spermatogonial stem cells

Yao¹,

Peng²,

Xu³

et al. 2020

Ann Transl Med

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Background: Spermatogonial stem cells and organ engineering research has raised new hope in infertility treatment. Spermatogenesis is a complex physiological process. To observe the proliferation ability and differentiation tendency of mice spermatogonial stem cells (SSCs), to study the effect of regulating the Wnt signaling pathway on the proliferation and differentiation of SSCs, and to provide a valuable basis for the clinical application of SSCs. Methods: SSCs were isolated and cultured by immunomagnetic separation. Cell surface markers were identified by flow cytometry. Axin1 was chosen as the target gene to inhibit fibrosis of SSCs by inhibiting the activity of Wnt signaling pathway. Axin-siRNA interference vector was constructed and transfected into spermatogonial stem cells. Cultured SSCs were randomly divided into six groups: control group, SSCs + TGF-β group, SSCs + DKK1 group, SSCs + Axin-RNAi group, SSCs + TGF-β + DKK1 group, SSCs + TGF-β + Axin-RNAi group. Proliferation of SSCs in each group was detected by MTT assay. Immunofluorescence, western blot and real time polymerase chain reaction analysis were used to detect protein expression in the Wnt/β catenin signaling pathways and the molecular markers of fibroblasts in SSCs.Results: Flow cytometry analysis confirmed that the cultured SSCs were of high purity. MTT assay showed there was no significant difference between Axin-siRNA transfected and non-transfected cells. The proliferation ability was significantly increased in the SSCs + TGF-β group, however, it was retarded in SSCs + Axin-RNAi group. The results of immunofluorescence and western blot analysis showed that the expression levels of the Wnt signaling pathway proteins were relatively inhibited after Axin-siRNA was applied. Real-time polymerase chain reaction showed that the expression levels of the molecular markers of fibroblasts were close to the normal control group. Conclusions:The Axin-siRNA constructed in this study specifically inhibited Wnt/β-catenin signal pathway activation, then inhibited the differentiation of SSCs into fibroblasts, which provides a valuable basis for the clinical application of SSCs.

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“…Stem cell transplantation has been explored as a therapy for various diseases such as stroke, 1 Alzheimer's disease, 2 diabetes mellitus, 3 Parkinson's, 4 myocardial infarction, 5 HIV, 6 testicular failure, 7,8 and other degenerative diseases such as ovarian failure. 9 As an important organ in female reproduction, the ovary produces ovum and female hormones.…”

Section: Introductionmentioning

confidence: 99%

Role of apoptotic inhibitors, viability, and differentiation in low oxygen tension of mesenchymal stem cells cultured in a rat model of ovarian failure

et al. 2023

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Background: Stem cell therapy shows applications potential for malnutrition-induced ovarian failure in rat models. However, it is ineffective because of the lack of viability and differentiation of transplanted stem cells, resulting in low adaptation and survival rates. We aimed to determine whether stem cells cultured under low oxygen (O2) tension improves the adaptability and viability of stem cells, as well as ovarian failure. Methods: After four days of culturing mesenchymal stem cells (MSCs) in 21% oxygen (normoxia) as the T2 group and 1% oxygen (low O2 or hypoxia) as the T1 group, 200 million bone marrow-derived MSCs per rat were transplanted into female rats with ovarian failure (15 rats per treatment group). A total of 15 fertile and 15 infertile rats were categorized as the C+ and C− groups, respectively. Results: The slight increase in cells expressing HSP70 (C+, T2, T1, and C− groups were 0.5a±0.53, 1.7a±0.82, 6.2b±1.5, and 9.6c±1.3, respectively), decrease in cells expressing caspase-3 as an apoptotic inhibitor (C+, T2, T1, and C− groups were 0.2a±0.42, 0.6a±0.52, 4.8b±1.03, and 7.3c±1.42, respectively), and increase in cells expressing VEGF-1 (C+, T2, T1, and C− groups were 10.8c±1.55, 8.7b±0.48, 0.4a±0.52, and 0.2a±0.42, respectively) and GDF-9 (C+, T2, T1, and C− groups were 5.8c±1.47, 4.6b±0.97, 0.5a±0.53, and 0.3a±0.48, respectively) were used as markers for viability and differentiation in ovarian tissue, indicating that MSCs cultured under low O2 tension were more effective than those cultured under normoxic conditions as a treatment for female rats with ovarian failure. Furthermore, infertile female rats treated with MSCs cultivated under low O2 tension had an enhanced ovarian tissue shape, as indicated by the increasing Graafian follicle count (C+, T2, T1, and C− groups were 8.9c±0.74, 4.5b±0.71, 0.5a±0.53, and 0.4a±0.52, respectively). Conclusions: MSCs cultured under low O2 tension are an effective treatment for malnourished rats with ovarian failure.

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Spermatogonial Stem Cell Transplantation in Large Animals

Cited by 15 publications

References 116 publications

Successful in vivo Transplantation of Cultured and Enriched Testicular Germ Cells of Pre-Pubertal Bucks to Busulfan-Treated Homologous Recipients

Successful in vivo Transplantation of Cultured and Enriched Testicular Germ Cells of Pre-Pubertal Bucks to Busulfan-Treated Homologous Recipients

The effect of regulating the Wnt signaling pathway on the proliferation and differentiation of spermatogonial stem cells

Role of apoptotic inhibitors, viability, and differentiation in low oxygen tension of mesenchymal stem cells cultured in a rat model of ovarian failure

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