In vitro complete differentiation of human spermatogonial stem cells to morphologic spermatozoa using a hybrid hydrogel of agarose and laminin

Jabari, Ayob; Gholami, Kheirollah; Khadivi, Farnaz; Koruji, Morteza; Amidi, Fardin; Gilani, Mohammad Ali Sadighi; Mahabadi, Vahid Pirhajati; Nikmahzar, Aghbibi; Salem, Maryam; Movassagh, Sepideh Ashouri; Feizollahi, Narjes; Abbasi, Mehdi

doi:10.1016/j.ijbiomac.2023.123801

Cited by 15 publications

(12 citation statements)

References 48 publications

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“…[36] Hydrogel-based models, on the other hand, involve embedding testicular cells within a 3D hydrogel matrix, creating a supportive environment for cell growth and differentiation. [35,37] These models provide a biomimetic microenvironment that promotes the viability and functionality of testicular cells. Although the developed scaffolds aimed to replicate the spatial characteristics of seminiferous tubules, there is room for improvement in mimicking the intricate architecture of native tissue.…”

Section: Discussionmentioning

confidence: 99%

Polycaprolactone/Testicular Extracellular Matrix/Graphene Oxide‐Based Electrospun Tubular Scaffolds for Reproductive Medicine: Biomimetic Architecture of Seminiferous Tubules

Mohammadi,

Koruji,

Azami

et al. 2023

Macromolecular Bioscience

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Numerous diverse scaffolds have been developed in the field of testicular bioengineering. However, effectively replicating the spatial characteristics of native tissue, poses a challenge in maintaining the requisite cellular arrangement essential for proper spermatogenesis. In order to mimic the structural properties of seminiferous tubules, our objective was to fabricate a biocompatible tubular scaffold. Following the decellularization process of the testicular tissue, validation of cellular remnants' elimination from the specimens was conducted using DAPI staining, H&E staining, and DNA content analysis. The presence of extracellular matrix components was confirmed through Alcian blue, Orcein, and Masson's trichrome staining techniques. The electrospinning technique was employed to synthesize the scaffolds using polycaprolactone, extracted ECM, and varying concentrations of graphene oxide (0.5%, 1%, and 2%). Subsequently, comprehensive evaluations were performed to assess the properties of the synthetic scaffolds. These evaluations encompassed Fourier‐transform infrared spectroscopy, scanning electron microscopy imaging, scaffold degradation testing, mechanical behavior analysis, MTT assay, and in vivo biocompatibility assessment. The PCL/dECM with 0.5% GO formulation exhibited superior fiber morphology, enhanced mechanical properties, and outperformed other groups in terms of in vitro biocompatibility. Consequently, these scaffolds present a viable option for implementation in “in vitro spermatogenesis” procedures, holding promise for future sperm production from spermatogonial cells.This article is protected by copyright. All rights reserved

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Section: Discussionmentioning

confidence: 99%

Polycaprolactone/Testicular Extracellular Matrix/Graphene Oxide‐Based Electrospun Tubular Scaffolds for Reproductive Medicine: Biomimetic Architecture of Seminiferous Tubules

Mohammadi,

Koruji,

Azami

et al. 2023

Macromolecular Bioscience

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“…For example, other components of the ECM (such as laminin and fibronectin) play an essential role in angiogenesis. [30,31] Therefore, modifying the surface of nanofibers with proteins similar to the physiological composition can be useful in creating a new vascular bed for reconstructive medical purposes. In the current study, hAM ECM proteins were extracted (as a human source not restricted in terms of ethics and regulation for preparing tissue-derived ECM) and their ability to promote cellular processes and tissue regeneration was compared with ECM derived from a animal specific tissue (bladder).…”

Section: Discussionmentioning

confidence: 99%

The Effect of Multilayered Electrospun PLLA Nanofibers Coated with Human Amnion or Bladder ECM Proteins on Epithelialization and Smooth Muscle Regeneration in the Rabbit Bladder

Gholami,

Seyedjafari,

Mahdavi

et al. 2023

Macromolecular Bioscience

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Nanofibrous scaffolds have attracted much attention in bladder reconstruction approaches due to their excellent mechanical properties. In addition, their biological properties can be improved by combination with biological materials. Taking into account the advantages of nanofibrous scaffolds and decellularized extracellular matrix (dECM) in tissue engineering, scaffolds of poly‑L‑lactic acid (PLLA) coated with decellularized human amnion membrane (hAM) or sheep bladder (SB)‐derived ECM proteins are developed (amECM‐coated PLLA and sbECM‐coated PLLA, respectively). The bladder regenerative potential of modified electrospun PLLA scaffolds is investigated in rabbits. The presence of ECM proteins is confirmed on the nanofibers’ surface. Coating the surface of the PLLA nanofibers improves cell adhesion and proliferation. Histological and immunohistochemical evaluations show that rabbits subjected to cystoplasty with a multilayered PLLA scaffold show de novo formation and maturation of the multilayered urothelial layer. However, smooth muscle bundles (myosin heavy chain [MHC] and α‐smooth muscle actin [α‐SMA] positive) are detected only in ECM‐coated PLLA groups. All groups show no evidence of a diverticulumor fistula in the urinary bladder. These results suggest that the biofunctionalization of electrospun PLLA nanofibers with ECM proteins can be a promising option for bladder tissue engineering. Furthermore, hAM can also replace animal‐sourced ECM proteins in bladder tissue regeneration approaches.

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“…In research published in 2012, mature mouse sperm were produced on a soft agar culture system (SACS) [99]. In another recent study, the completion of human spermatogenesis was observed on agarose gel plus a laminin supplement in the presence of Sertoli cells after being cultured for 74 days [100]. Analysis showed that the laminin-enhanced 3D matrix supported all physiological activities of the SSCs, including survival and proliferation, and led to the differentiation of spermatogonial cells into morphological sperm.…”

Section: Ssc Culturementioning

confidence: 99%

In vivo and in vitro sperm production: an overview of the challenges and advances in male fertility restoration

Bashiri,

Hosseini,

Salem

et al. 2024

Clin Exp Reprod Med

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Male infertility can be caused by genetic anomalies, endocrine disorders, inflammation, and exposure to toxic chemicals or gonadotoxic treatments. Therefore, several recent studies have concentrated on the preservation and restoration of fertility to enhance the quality of life for affected individuals. It is currently recommended to biobank the tissue extracted from testicular biopsies to provide a later source of spermatogonial stem cells (SSCs). Another successful approach has been the in vitro production of haploid male germ cells. The capacity of SSCs to transform into sperm, as in testicular tissue transplantation, SSC therapy, and in vitro or ex vivo spermatogenesis, makes them ideal candidates for in vivo fertility restoration. The transplantation of SSCs or testicular tissue to regenerate spermatogenesis and create embryos has been achieved in nonhuman mammal species. Although the outcomes of human trials have yet to be released, this method may soon be approved for clinical use in humans. Furthermore, regenerative medicine techniques that develop tissue or cells on organic or synthetic scaffolds enriched with bioactive molecules have also gained traction. All of these methods are now in different stages of experimentation and clinical trials. However, thanks to rigorous studies on the safety and effectiveness of SSC-based reproductive treatments, some of these techniques may be clinically available in upcoming decades.

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In vitro complete differentiation of human spermatogonial stem cells to morphologic spermatozoa using a hybrid hydrogel of agarose and laminin

Cited by 15 publications

References 48 publications

Polycaprolactone/Testicular Extracellular Matrix/Graphene Oxide‐Based Electrospun Tubular Scaffolds for Reproductive Medicine: Biomimetic Architecture of Seminiferous Tubules

Polycaprolactone/Testicular Extracellular Matrix/Graphene Oxide‐Based Electrospun Tubular Scaffolds for Reproductive Medicine: Biomimetic Architecture of Seminiferous Tubules

The Effect of Multilayered Electrospun PLLA Nanofibers Coated with Human Amnion or Bladder ECM Proteins on Epithelialization and Smooth Muscle Regeneration in the Rabbit Bladder

In vivo and in vitro sperm production: an overview of the challenges and advances in male fertility restoration

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