Evaluation of co-cultured spermatogonial stem cells encapsulated in alginate hydrogel with Sertoli cells and their transplantation into azoospermic mice

Veisi, Mohammad; Mansouri, Kamran; Assadollahi, Vahideh; Jalili, Cyrus; Pirnia, Afshin; Salahshoor, Mohammad Reza; Hoseinkhani, Zohreh; Gholami, Mohammad

doi:10.1017/s0967199421000733

Cited by 11 publications

(10 citation statements)

References 40 publications

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“…We cultured human testicular cells on human DTM for the proliferation and differentiation of SSCs for 4 weeks. Long‐term SCCs culture can lead to the loss of specific properties and induce apoptosis in these cells 34 . Thus creating an efficient 3D culturing system can be essential in SSCs maintenance, proliferation, and differentiation.…”

Section: Discussionmentioning

confidence: 99%

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Differentiation of human spermatogonial stem cells using a human decellularized testicular scaffold supplemented by platelet‐rich plasma

et al. 2023

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Background: Effective culture systems for attachment, migration, proliferation, and differentiation of spermatogonial stem cells (SSCs) can be a promising therapeutic modality for preserving male fertility. Decellularized extracellular matrix (ECM) from native testis tissue creates a local microenvironment for testicular cell culture. Furthermore, platelet-rich plasma (PRP) contains various growth factors for the proliferation and differentiation of SSCs. Methods:In this study, human testicular cells were isolated and cultured for 4 weeks, and SSCs were characterized using immunocytochemistry (ICC) and flow cytometry. Human testicular tissue was decellularized (0.3% SDS, 1% Triton), and the efficiency of the decellularization process was confirmed by histological staining and DNA content analysis. SSCs were cultured on the human decellularized testicular matrix (DTM) for 4 weeks. The viability and the expression of differentiation genes were evaluated by MTT and real-time polymerase chain reaction (PCR), respectively.Results: Histological evaluation and DNA content analysis showed that the components of ECM were preserved during decellularization. Our results showed that after 4 weeks of culture, the expression levels of BAX, BCL-2, PLZF, and SCP3 were unchanged, while the expression of PRM2 significantly increased in the cells cultured on DTM supplemented with PRP (ECM-PRP). In addition, the expression of GFRA1 was significantly decreased in the ECM group compared to the control and PRP groups. Furthermore, the MTT test indicated that viability was significantly enhanced in cells plated on DTM supplemented with PRP. Conclusion:Our study demonstrated that DTM supplemented with PRP can provide an effective culture system for the differentiation and viability of SSCs.

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Section: Discussionmentioning

confidence: 99%

“…This finding can originate from the important role of testis ECM components together with the PRP factors affecting SSCs viability and differentiation 3,29,36,40,43 . Apoptosis in SSCs can be induced in long‐term cell culture 34 . So, we evaluated the expression level of BCL‐2 and BAX after 4 weeks of culture.…”

Section: Discussionmentioning

confidence: 99%

Differentiation of human spermatogonial stem cells using a human decellularized testicular scaffold supplemented by platelet‐rich plasma

et al. 2023

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“…Veisi et al. (2022) ( 239 ) used co-cultured spermatogonial stem cells encapsulated in alginate hydrogel with Sertoli cells and found that culturing SSCs in alginate hydrogel with Sertoli cells in a 3D culture can lead to efficient proliferation and maintenance of SSC and increase the efficiency of SSC transplantation. Poels et al.…”

Section: Discussionmentioning

confidence: 99%

Biomaterials for Testicular Bioengineering: How far have we come and where do we have to go?

et al. 2023

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Traditional therapeutic interventions aim to restore male fertile potential or preserve sperm viability in severe cases, such as semen cryopreservation, testicular tissue, germ cell transplantation and testicular graft. However, these techniques demonstrate several methodological, clinical, and biological limitations, that impact in their results. In this scenario, reproductive medicine has sought biotechnological alternatives applied for infertility treatment, or to improve gamete preservation and thus increase reproductive rates in vitro and in vivo. One of the main approaches employed is the biomimetic testicular tissue reconstruction, which uses tissue-engineering principles and methodologies. This strategy pursues to mimic the testicular microenvironment, simulating physiological conditions. Such approach allows male gametes maintenance in culture or produce viable grafts that can be transplanted and restore reproductive functions. In this context, the application of several biomaterials have been proposed to be used in artificial biological systems. From synthetic polymers to decellularized matrixes, each biomaterial has advantages and disadvantages regarding its application in cell culture and tissue reconstruction. Therefore, the present review aims to list the progress that has been made and the continued challenges facing testicular regenerative medicine and the preservation of male reproductive capacity, based on the development of tissue bioengineering approaches for testicular tissue microenvironment reconstruction.

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“…The 3D decellularized testicular scaffold with hyaluronic acid and chitosan provides the condition for the differentiation and proliferation of mice SSCs ( 102 ). The proliferation and self-renewal of mice SSCs was stimulated by culturing on the 3D scaffold consisting of alginate hydrogel with Sertoli cells ( 103 ). The mice germ cells were cultured in 3D printed one-layer scaffolds at the air-medium interface simulating the tubule-like structure.…”

Section: Fertility Restoration Through In Vitro Sp...mentioning

confidence: 99%

Roles of Spermatogonial Stem Cells in Spermatogenesis and Fertility Restoration

et al. 2022

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Spermatogonial stem cells (SSCs) are a group of adult stem cells in the testis that serve as the foundation of continuous spermatogenesis and male fertility. SSCs are capable of self-renewal to maintain the stability of the stem cell pool and differentiation to produce mature spermatozoa. Dysfunction of SSCs leads to male infertility. Therefore, dissection of the regulatory network of SSCs is of great significance in understanding the fundamental molecular mechanisms of spermatogonial stem cell function in spermatogenesis and the pathogenesis of male infertility. Furthermore, a better understanding of SSC biology will allow us to culture and differentiate SSCs in vitro, which may provide novel stem cell-based therapy for assisted reproduction. This review summarizes the latest research progress on the regulation of SSCs, and the potential application of SSCs for fertility restoration through in vivo and in vitro spermatogenesis. We anticipate that the knowledge gained will advance the application of SSCs to improve male fertility. Furthermore, in vitro spermatogenesis from SSCs sets the stage for the production of SSCs from induced pluripotent stem cells (iPSCs) and subsequent spermatogenesis.

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Evaluation of co-cultured spermatogonial stem cells encapsulated in alginate hydrogel with Sertoli cells and their transplantation into azoospermic mice

Cited by 11 publications

References 40 publications

Differentiation of human spermatogonial stem cells using a human decellularized testicular scaffold supplemented by platelet‐rich plasma

Differentiation of human spermatogonial stem cells using a human decellularized testicular scaffold supplemented by platelet‐rich plasma

Biomaterials for Testicular Bioengineering: How far have we come and where do we have to go?

Roles of Spermatogonial Stem Cells in Spermatogenesis and Fertility Restoration

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