Meiotic activity in orthotopic xenografts derived from human postpubertal testicular tissue

Saen, Dorien Van; Goossens, Ellen; Bourgain, Claire; Ferster, Alina; Tournaye, Herman

doi:10.1093/humrep/deq321

Cited by 78 publications

(61 citation statements)

References 32 publications

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“…Such intact clusters (ICs) are seen as a small or larger group of LCs in close contact with the surrounding connective tissue and in the vicinity or around blood vessels (Davidoff et al, 2009). Staining procedures for inhibin a and StAR were performed as previously described (Van Saen et al, 2011).…”

Section: Light Microscopymentioning

confidence: 99%

What is the best cryopreservation protocol for human testicular tissue banking?

et al. 2013

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study question: Is there a better alternative to the conventional cryopreservation protocols for human testicular tissue banking? summary answer: Uncontrolled slow freezing (USF) using 1.5 M dimethylsulphoxide (DMSO) and 0.15 M sucrose as cryoprotectants appears to be a user-friendly and efficient method for the cryopreservation of human testicular tissue.what is known already: Currently, time-consuming controlled slow freezing (CSF) protocols that need expensive equipment are commonly used for human testicular tissue banking. USF and vitrification are cryopreservation techniques that were successfully applied in several animal models but need further exploration with human tissue. study design, size, duration: Fragments (n ¼ 160) of testicular tissue from 14 patients undergoing vasectomy reversal were assigned to a fresh control group or one of the following cryopreservation procedures: CSF using DMSO at a concentration of 0.7 or 1.5 M in the presence (+S) or absence of sucrose (2S), USF using either 0.7 or 1.5 M DMSO combined with sucrose, solid-surface vitrification (SSV) or direct cover vitrification (DCV). materials, setting, methods: Light microscopic evaluations were performed to study apoptosis, germ cell proliferation ability, spermatogonial survival, coherence of the seminiferous epithelium and integrity of the interstitial compartment after cryopreservation. Ultrastructural alterations were studied by scoring cryodamage to four relevant testicular cell types.main results and the role of chance: The USF 1.5 M DMSO + S protocol proved not solely to prevent cell death and to preserve seminiferous epithelial coherence, interstitial compartment integrity, SG and their potential to divide but also protected the testicular cell ultrastructure. A significant reduction in the number of SG per tubule from 21.4 + 5.6 in control tissue to 4.9 + 2.1, 8.2 + 5.4, 11.6 + 5.1, 8.8 + 3.9, 12.6 + 4.4 and 11.7 + 5.7 was observed after cryopreservation combined with at least one other form of cryoinjury when using CSF 0.7 M DMSO 2S, CSF 0.7 M DMSO + S, CSF 1.5 M DMSO + S, USF 0.7 M DMSO + S, SSV and direct cover vitrification (DCV), respectively (P , 0.001).limitations, reasons for caution: Supplementary research is required to investigate the effect on tissue functionality and to confirm this study's findings using prepubertal tissue.wider implications of the findings: An optimal cryopreservation protocol enhances the chances for successful fertility restoration. USF, being an easy and cost-effective alternative to CSF, would be preferable for laboratories in developing countries or whenever tissue is to be procured from a diseased child at a site distant from the banking facility.

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Section: Light Microscopymentioning

confidence: 99%

What is the best cryopreservation protocol for human testicular tissue banking?

et al. 2013

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“…The testicular biopsies were fixed and embedded in paraffin at the pathology department of the Universitair Ziekenhuis Brussel. Five-micrometer-thick sections were stained with hematoxylin and eosin to evaluate structural integrity, and immunohistochemical stainings were performed for melanoma-associated antigen 4 (MAGE-A4; provided by Dr. Giulio Spagnoli, University of Basel, Switzerland) and vimentin (mouse monoclonal antibody clone V9, M0725, Dako) as previously described (18). The seminiferous tubules were classified as normal (intact basement membrane, good attachment of cells to the basement membrane, and intercellular adhesion), degenerated (degenerative Sertoli cells with pyknotic nuclei, detachment of cells from the basement membrane, the loss of intercellular contacts, and moderate thickened basal lamina), and hyalinized (massive thickening of the basement membrane and complete loss of seminiferous epithelial cells).…”

Section: Histologymentioning

confidence: 99%

Presence of spermatogonia in 47,XXY men with no spermatozoa recovered after testicular sperm extraction

Saen

Tournaye

Goossens

2012

Fertility and Sterility

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“…The cryopreserved testicular tissue may also serve as an autologous source of pluripotent stem cells for future regenerative medicine (5)(6)(7)(8)(9). Although ovarian cryopreservation has successfully produced live human births (10,11), recapitulation of stages during human spermatogenesis after freezing is still elusive (12)(13)(14). Continued efforts directed toward optimization of cryopreservation protocols are necessary for improvement of postthaw testicular cell functionality.…”

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confidence: 99%