Cryostorage of testicular tissue and retransplantation of spermatogonial stem cells in the infertile male

Kanbar, Marc; Michele, Francesca de; Wyns, Christine

doi:10.1016/j.beem.2018.10.003

Cited by 18 publications

(8 citation statements)

References 87 publications

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“…This procedure would not allow to preserve the spermatogenetic niche in its entirety ( 95 , 96 ) and has also been studied in humans less intensively ( 97 ). Freezing cells rather than tissue fragments will make it impossible to employ some techniques, such as testicular graft transplantation or tissue culture, whereas a cryopreserved tissue fragment could undergo further enzymatic digestion to obtain SSCs and other testicular cells ( 98 ). It is essential to improve the freezing protocol, trying to reduce the generation of alterations in thawed sperm quality ( 99 ).…”

Section: Cryopreservation: Testicular Tissue or Cell Suspensionmentioning

confidence: 99%

Fertility Preservation and Restoration Options for Pre-Pubertal Male Cancer Patients: Current Approaches

et al. 2022

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Fertility preservation for prepubertal male patients undergoing gonadotoxic therapies, potentially depleting spermatogonial cells, is an expanding necessity, yet most of the feasible options are still in the experimental phase. We present our experience and a summary of current and novel possibilities regarding the different strategies to protect or restore fertility in young male patients, before proceeding with chemotherapy or radiotherapy for malignances or other diseases. Adult oncological patients should always be counselled to cryopreserve the semen before starting treatment, however this approach is not suitable for prepubertal boys, who aren’t capable to produce sperm yet. Fortunately, since the survival rate of pediatric cancer patients has skyrocketed in the last decade and it’s over 84%, safeguarding their future fertility is becoming a major concern for reproductive medicine. Surgical and medical approaches to personalize treatment or protect the gonads could be a valid first step to take. Testicular tissue autologous grafting or xenografting, and spermatogonial stem cells (SSCs) transplantation, are the main experimental options available, but spermatogenesis in vitro is becoming an intriguing alternative. All of these methods feature both strong and weak prospects. There is also relevant controversy regarding the type of testicular material to preserve and the cryopreservation methods. Since transplanted cells are bound to survive based on SSCs number, many ways to enrich their population in cultures have been proposed, as well as different sites of injection inside the testis. Testicular tissue graft has been experimented on mice, rabbits, rhesus macaques and porcine, allowing the birth of live offspring after performing intracytoplasmic sperm injection (ICSI), however it has never been performed on human males yet. In vitro spermatogenesis remains a mirage, although many steps in the right direction have been performed. The manufacturing of 3D scaffolds and artificial spermatogenetic niche, providing support to stem cells in cultures, seems like the best way to further advance in this field.

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Section: Cryopreservation: Testicular Tissue or Cell Suspensionmentioning

confidence: 99%

Fertility Preservation and Restoration Options for Pre-Pubertal Male Cancer Patients: Current Approaches

et al. 2022

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“…These biophysical transfer phenomena are substantially reduced when using microscopic SSC suspensions. One major advantage to TT, however, is preservation of the SSC niche architecture, which may improve post-thaw SSC viability and function (67).…”

Section: Testicular Tissue Versus Cell Suspensionmentioning

confidence: 99%

“…Therefore, the rarity of SSCs may be less of a problem for humans than rodents. Nonetheless, it is reasonable to expect that increasing the number of transplanted SSCs correlates with more robust engraftment and regeneration of spermatogenesis (67,88).…”

Section: Restoring Sperm Production Autologous Approachesmentioning

confidence: 99%

Progress in translational reproductive science: testicular tissue transplantation and in vitro spermatogenesis

Pelzman

Orwig

Hwang

2020

Fertility and Sterility

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Since the birth of the first child conceived via in vitro fertilization 40 years ago, fertility treatments and assisted reproductive technology have allowed many couples to reach their reproductive goals. As of yet, no fertility options are available for men who cannot produce functional sperm, but many experimental therapies have demonstrated promising results in animal models. Both autologous (stem cell transplantation, de novo morphogenesis, and testicular tissue grafting) and outside-the-body (xenografting and in vitro spermatogenesis) approaches exist for restoring sperm production in infertile animals with varying degrees of success. Once safety profiles are established and an ideal patient population is chosen, some of these techniques may be ready for human experimentation in the near future, with likely clinical implementation within the next decade. (Fertil Steril Ò 2020;113:500-9. Ó2020 by American Society for Reproductive Medicine.

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“…10 In vitro maturation (IVM) of spermatogonial stem cells obtained from cryopreserved pre-pubertal testicular tissues via two-dimensional (2D) and three-dimensional (3D) culture systems is the best potential procedure for the generation of functional sperm. 11,12 2D culture systems cannot effectively simulate the organized structure of the testicular microenvironment. In contrast, a 3D culture system, such as organotypic or organoid culture, can provide an ideal environment similar to the SSCs niches in seminiferous tubules.…”

Section: Introductionmentioning

confidence: 99%

Differentiation of human primary testicular cells in the presence of SCF using the organoid culture system

Nikmahzar,

Koruji,

Jahanshahi

et al. 2023

Artificial Organs

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PurposeDevelopment of organoids using human primary testicular cells has remained a challenge due to the complexity of the mammalian testicular cytoarchitecture and culture methods. In this study, we generated testicular organoids derived from human primary testicular cells. Then, we evaluated the effect of stem cell factor (SCF) on cell differentiation and apoptosis in the testicular organoid model.MethodsThe testicular cells were harvested from the three brain‐dead donors. Human spermatogonial stem cells (SSCs) were characterized using immunocytochemistry (ICC), RT‐PCR and flow cytometry. Testicular organoids were generated from primary testicular cells by hanging drop culture method and were cultured in three groups: control group, experimental group 1 (treated FSH and retinoic acid (RA)), and experimental group 2 (treated FSH, RA and SCF), for five weeks. We assessed the expression of SCP3 (Synaptonemal Complex Protein 3) as a meiotic gene, PRM2 (Protamine 2) as a post‐meiotic marker and apoptotic genes of Bax (BCL2‐Associated X Protein) and Bcl‐2 (B‐cell lymphoma 2), respectively by using RT‐qPCR. In addition, we identified the expression of PRM2 by immunohistochemistry (IHC).ResultsRelative expression of SCP3, PRM2 and Bcl‐2 were highest in group 2 after five weeks of culture. In contrast, BAX expression level was lower in experimental group 2 in comparison with other groups. IHC analyses indicated the highest expression of PRM2 as a postmeiotic marker in group 2 in comparison to 2D culture and control groups but not find significant differences between experimental group 1 and experimental group 2 groups. Morphological evaluations revealed that organoids are compact spherical structures and in the peripheral region composed of uncharacterized elongated fibroblast‐like cells.ConclusionOur findings revealed that the testicular organoid culture system promote the spermatogonial stem cell (SSC) differentiation, especially in presence of SCF. Developed organoids are capable of recapitulating many important properties of a stem cell niche.

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Cryostorage of testicular tissue and retransplantation of spermatogonial stem cells in the infertile male

Cited by 18 publications

References 87 publications

Fertility Preservation and Restoration Options for Pre-Pubertal Male Cancer Patients: Current Approaches

Fertility Preservation and Restoration Options for Pre-Pubertal Male Cancer Patients: Current Approaches

Progress in translational reproductive science: testicular tissue transplantation and in vitro spermatogenesis

Differentiation of human primary testicular cells in the presence of SCF using the organoid culture system

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