Differentiation and proliferation of spermatogonial stem cells using a three-dimensional decellularized testicular scaffold: a new method to study the testicular microenvironment in vitro

Introduction: Induction of in vitro spermatogenesis can be useful for infertility treatment in azoospermic patients and those undergoing chemotherapy. Different culture systems have been used to achieve this goal. This review study was performed with the aim to evaluate the application of 3D culture and testicular scaffolds in the establishment of in vitro spermatogenesis. In this review study, the information on the application of 3D culture and testicular scaffolds in induction of in vitro spermatogenesis was searched in databases such as SID, Magiran, PubMed, Irandoc, Iranmedx Scopus, Google Scholar, Web of Science using the keywords of three dimensional culture, testicular scaffold, spermatogenesis, spermatogonial stem cells without time limitation. Data analysis was carried out qualitatively. Finally, 35 papers in English and Persian were used to compile the article. In order to induce of in vitro spermatogenesis, three-dimensional culture methods such as testicular tissue culture, soft agar culture system, natural biomaterial scaffolds such as collagen, and scaffolds derived from decellularized testis have been used. Conclusion: Three-dimensional culture using spermatogonial stem cells and scaffolds can be used in vitro for induction of spermatogenesis, but there are further technical and ethical challenges in the path of fertile sperm production for the treatment of infertility.

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IN VITRO DIFFERENTIATION OF HUMAN SPERMATOGONIAL STEM CELLS IN A THREE DIMENSIONAL (3D) TESTICULAR ORGANOID SYSTEM

The Journal of Urology ◽

10.1016/j.juro.2017.02.1287 ◽

2017 ◽

Vol 197 (4S) ◽

Cited By ~ 1

Author(s):

Kara E. McAbee ◽

Nima Pourhabibi Zarandi ◽

Anthony Atala ◽

Hooman Sadri-Ardekani ◽

Colin Bishop

Keyword(s):

Stem Cells ◽

Three Dimensional ◽

Spermatogonial Stem Cells ◽

In Vitro Differentiation

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In Vitro Spermatogenesis by Three-dimensional Culture of Spermatogonial Stem Cells on Decellularized Testicular Matrix

10.31661/gmj.v8i0.1565 ◽

2019 ◽

Vol 8 ◽

pp. 1565 ◽

Cited By ~ 1

Author(s):

Sepideh Ashouri Movassagh ◽

Mehdi Banitalebi Dehkordi ◽

Morteza Koruji ◽

Gholamreza Pourmand ◽

Parvaneh Farzaneh ◽

...

Keyword(s):

Stem Cells ◽

Male Infertility ◽

Culture Medium ◽

Three Dimensional ◽

Cell Interaction ◽

Spermatogonial Stem Cells ◽

3D Matrix ◽

Specific Culture

Background: In the males, Spermatogonial Stem Cells (SSCs) contribute to the production of sex cells and fertility. In vitro SSCs culture can operate as an effective strategy for studies on spermatogenesis and male infertility treatment. Cell culture in a three-dimensional (3D) substrate, relative to a two-dimensional substrate (2D), creates better conditions for cell interaction and is closer to in vivo conditions. In the present study, in order to create a 3D matrix substrate, decellularized testicular matrix (DTM) was used to engender optimal conditions for SSCs culture and differentiation. Materials and Methods: After, testicular cells enzymatic extraction from testes of brain-dead donors, the SSCs were proliferated in a specific culture medium for four weeks, and after confirming the identity of the colonies derived from the growth of these cells, they were cultured on a layer of DTM as well as in 2D condition with a differentiated culture medium. In the Sixth week since the initiation of the differentiation culture, the expression of pre meiotic (OCT4 & PLZF), meiotic (SCP3 & BOULE) and post meiotic (CREM & Protamine-2) genes were measured in both groups. Results: The results indicated that the expression of pre meiotic, meiotic and post meiotic genes was significantly higher in the cells cultured on DTM (P ≤ 0.001). Conclusion: SSCs culture in DTM with the creation of ECM and similar conditions with in vivo can be regarded as a way of demonstrating spermatogenesis in vitro, which can be adopted as a treatment modality for male infertility. [GMJ.2019;8:e1565]

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In vitro differentiation of spermatogonial stem cells to functional sperm in zebrafish

Reproduction Abstracts ◽

10.1530/repabs.1.p356 ◽

2014 ◽

Author(s):

Noriyoshi Sakai

Keyword(s):

Stem Cells ◽

Spermatogonial Stem Cells ◽

In Vitro Differentiation

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Homing of mouse spermatogonial stem cells on acellular testis following in vitro transplantation.

10.26226/morressier.5af300b3738ab10027aa9b14 ◽

2018 ◽

Author(s):

Mansoureh Movahedin

Keyword(s):

Stem Cells ◽

Spermatogonial Stem Cells

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In Vitro Propagation of Human Prepubertal Spermatogonial Stem Cells

JAMA ◽

10.1001/jama.2011.791 ◽

2011 ◽

Vol 305 (23) ◽

pp. 2416 ◽

Cited By ~ 139

Author(s):

Hooman Sadri-Ardekani

Keyword(s):

Stem Cells ◽

In Vitro Propagation ◽

Spermatogonial Stem Cells

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Propagation of bovine spermatogonial stem cells in vitro

Reproduction ◽

10.1530/rep-07-0419 ◽

2008 ◽

Vol 136 (5) ◽

pp. 543-557 ◽

Cited By ~ 90

Author(s):

Pedro M Aponte ◽

Takeshi Soda ◽

Katja J Teerds ◽

S Canan Mizrak ◽

Henk J G van de Kant ◽

...

Keyword(s):

Stem Cells ◽

Growth Factor ◽

Cultured Cells ◽

Somatic Cells ◽

Type A ◽

Spermatogonial Stem Cells ◽

Seminiferous Tubules ◽

Type A Spermatogonia ◽

Stimulation Of

The access to sufficient numbers of spermatogonial stem cells (SSCs) is a prerequisite for the study of their regulation and further biomanipulation. A specialized medium and several growth factors were tested to study thein vitrobehavior of bovine type A spermatogonia, a cell population that includes the SSCs and can be specifically stained for the lectin Dolichos biflorus agglutinin. During short-term culture (2 weeks), colonies appeared, the morphology of which varied with the specific growth factor(s) added. Whenever the stem cell medium was used, round structures reminiscent of sectioned seminiferous tubules appeared in the core of the colonies. Remarkably, these round structures always contained type A spermatogonia. When leukemia inhibitory factor (LIF), epidermal growth factor (EGF), or fibroblast growth factor 2 (FGF2) were added, specific effects on the numbers and arrangement of somatic cells were observed. However, the number of type A spermatogonia was significantly higher in cultures to which glial cell line-derived neurotrophic factor (GDNF) was added and highest when GDNF, LIF, EGF, and FGF2 were all present. The latter suggests that a proper stimulation of the somatic cells is necessary for optimal stimulation of the germ cells in culture. Somatic cells present in the colonies included Sertoli cells, peritubular myoid cells, and a few Leydig cells. A transplantation experiment, using nude mice, showed the presence of SSCs among the cultured cells and in addition strongly suggested a more than 10 000-fold increase in the number of SSCs after 30 days of culture. These results demonstrate that bovine SSC self-renew in our specialized bovine culture system and that this system can be used for the propagation of these cells.

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