Orthotopic grafting of cryopreserved prepubertal testicular tissue: in search of a simple yet effective cryopreservation protocol

Baert, Yoni; Goossens, Ellen; Saen, Dorien Van; Liang, Ning; Veld, Peter In’t; Tournaye, Herman

doi:10.1016/j.fertnstert.2012.02.010

Cited by 60 publications

(82 citation statements)

References 21 publications

(28 reference statements)

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“…Cell viability after thawing was higher in the vitrification group (95%) and rapid freezing group (72%) as compared to the slow freezing group (24%). In another study, conducted by BAERT et al (2012), testicular tissue fragments of These results are mostly consistent with those reported by THUWANUT et al (2013). They assessed the effect of slow and rapid freezing on the testicular tissue of wild cats (Felis chaus; Panthera leo; Panthera pardus), and reported that testicular tissue subjected to rapid freezing had better plasma membrane integrity.…”

Section: Cryopreservation Methods Of Testicular Tissuesupporting

confidence: 78%

“…Final CPA concentration is high, at more than 30% (ABRISHAMI et al, 2010, NOHALEZ et al, 2015. The solution containing CPAs is added to cell culture media, along with external CPAs such as sucrose and fetal bovine serum (BAERT et al, 2012). After the testicular tissue is exposed to the vitrification solution, fragments are placed in a metal cube held above liquid nitrogen in an insulated box, which enables the tissue to be cooled ultrarapidly.…”

Section: Cryopreservation Methods Of Testicular Tissuementioning

confidence: 99%

“…It is also more efficient than other techniques in avoiding crystallization due to the ultra-fast cooling rates used. However, exposing tissue to high concentrations of CPAs can be rather detrimental to the maintenance of morphological and functional characteristics of stem cells (BAERT et al, 2012).…”

Section: Cryopreservation Methods Of Testicular Tissuementioning

confidence: 99%

“…One extracellular CPA used in research involving testicular cryopreservation is sucrose, which is always associated with intracellular CPAs in cryoprotectant addition protocols (BAERT et al, 2012).…”

Section: Cryoprotectant Agentsmentioning

confidence: 99%

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Cryopreservation of testicular tissue: an alternative to maintain the reproductive capacity in different animal species

Lima

Silva

2017

Cienc. Rural

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Cryopreservation of testicular tissue enables the maintenance of reproductive capacity in different animal species, and contributes to the formation of gene banks for endangered species. The spermatogonia present in the testes can be grown in vitro and the sperm obtained can be used in artificial breeding programs. This review aimed to describe the main techniques of testicular cryopreservation, the main cryoprotectants used, as well as the progress made in different animal species thus far. In the last decade, significant progress has been made in obtaining viable and functional germ cells from testicular tissue. However, more research is needed to better establish protocols that can be used in clinical practice with various species.

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Section: Cryopreservation Methods Of Testicular Tissuesupporting

confidence: 78%

Section: Cryopreservation Methods Of Testicular Tissuementioning

confidence: 99%

Section: Cryopreservation Methods Of Testicular Tissuementioning

confidence: 99%

Section: Cryoprotectant Agentsmentioning

confidence: 99%

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Cryopreservation of testicular tissue: an alternative to maintain the reproductive capacity in different animal species

Lima

Silva

2017

Cienc. Rural

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“…Endoplazmik retikulumların (ER) şişip genişlemesi, mitokondrilerde yoğun görünüm, nukleuslarda hafif ve orta derecede heterokromatin yapısının görülmesi geri dönüşümlü (reversible) değişiklikler olarak kabul edildi. Mitokondrilerde atılmış yün görünümü, ER'de aşırı şişme genişlemeyle birlikte sitoplazmada geniş vakuollerin oluşması, nukleuslarda piknoz, nukleus membranlarında ayrışmalar ve ondülasyonlar geri dönüşümsüz (irreversible) değişiklikler olarak kabul edildi [23,24].…”

Section: Kesitlerin Histolojik Değerlendirilmesiunclassified

Prepubertal testis hücre süspansiyonun dondurulmasında seeding etkisi

Kervancıoğlu¹,

Çetınel²,

Demirci³

et al. 2015

Marmara Medical Journal

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ÖZETAmaç: Prepubertal testis kriyoprezervasyonu, kanser tedavisi yapılacak erkek çocuklarda fertilitenin korunması için günümüz teknolojisinde umut vadetmektedir. Kriyoprezervasyonda en uygun yöntemin geliştirilmesi için prepubertal testis hücre süspansiyonunda farklı taşıyıcılar kullanarak seeding etkisi araştırıldı. Gereç ve Yöntem:Yedi-sekiz günlük Wistar yavru erkek sıçan testis hücre süspansiyonları, programlı yavaş dondurma yöntemiyle donduruldu. Taşıyıcı olarak seçilen yüksek güvenlikli straw ve kriyotüpe seeding uygulandı. Spermatogonyal kök hücreler ve Sertoli hücreleri üzerindeki morfolojik etki, semi-kantitatif olarak ışık, kalitatif olarak geçirimli elektron mikroskobunda incelendi. Bulgular:Canlılık oranı, yüksek güvenlikli straw'da seeding uygulanmayan ve uygulanan gruplarda sırasıyla %90,6±3,0, %89,4±1,3, kriyotüpte %77,6±4,2 ve %85,4±2,7 bulundu. Seeding uygulanmasıyla, canlılık oranında straw grupları arasında fark görülmemesine rağmen, kriyotüp grupları arasında fark anlamlıydı (p<0,0001). Straw1. ve 2. grupları ile kriyotüp 3. ve 4. grupları arasında da fark anlamlıydı (p<0,0001 ve p< 0,05). Spermatogonyal kök hücreler ve Sertoli hücrelerinin ışık ve elektron mikroskobik incelemesinde, nukleus ve sitoplazmik yapıların ve membranların seeding uygulanan gruplarda belirgin fark yarattığı, ayrıca yüksek güvenlikli straw'ın testis hücre süspansiyonu için kriyotüpe oranla daha üstün bir taşıyıcı olduğu görüldü. Sonuç:Prepubertal testis hücre süspansiyonunun kriyoprezervasyonunda farklı taşıyıcılarda seeding etkisinin ilk defa kıyaslandığı çalışmamızda, taşıyıcı olarak yüksek güvenlikli straw'ın kullanıldığı seeding uygulanan yöntemin en uygun yöntem olduğu sonucuna varıldı.Anahtar kelimeler: Prepubertal testis, Testis hücre süspansiyonu, Programlı yavaş dondurma, Spermatogonyal kök hücre, Seeding ABSTRACT Objective: The cryopreservation of prepubertal testicular tissue promises to preserve fertility in boys under cancer treatment. The effects of a seeding process was investigated using different carriers for prepubertal testicular cell suspensions in order to develop an appropriate method for cryopreservation. Materials and Methods:Testicular cell suspensions of 7-8 day old male Wistar rats were frozen by a programmed slow freezing method. The straw and cryotube selected as carriers underwent a seeding process. The morphological effects on spermatogonial stem cells and Sertoli cells were evaluated semi-quantitatively by light microscopy and qualitatively with a transmission electron microscope. Results:The rate of vitality with or without seeding in straw was respectively, 90.6% (±3.0), 89.4 (±1.3) and in cryotube 77.6 (±4.2) and 85.4 (±2.7). The seeding process, showed no significant difference between the straw groups, but a significant difference was seen between the cryotube groups (p<0.0001). The nuclei, cytoplasmic structures and membranes of Sertoli and spermatogonial stem cells, showed less degeneration in the straw than in the cryotubes when the seeding process was applied. Conclusion:In th...

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Optimizing methods for human testicular tissue cryopreservation and spermatogonial stem cell isolation

Moraveji

Esfandiari

Sharbatoghli

et al. 2018

J of Cellular Biochemistry

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Cryopreservation of testicular tissue before cancer therapy for fertility preservation in prepubertal boys with cancer is of great interest in reproductive medicine. Isolation of spermatogonial stem cells (SSCs) from cryopreserved tissues would be a suitable cell source to re‐establish spermatogenesis after cancer therapy. We herein establish optimized protocols for cryopreservation of human testicular tissue and isolation of SSCs from cryopreserved tissue. We developed a freezing protocol that provided high testicular cell viability and supported structural integrity and tubular epithelium coherence similar to fresh tissue. Then, we established a protocol that allowed efficient isolation of functional SSCs from cryopreserved tissues. Isolated cells were found on the testicular basement membrane after xenotransplantation. Our results demonstrated the preservation of testicular tissue structure and high cell viability with efficient isolation of SSCs after testicular cryopreservation, which is promising for future therapeutic applications in fertility preservation.

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Orthotopic grafting of cryopreserved prepubertal testicular tissue: in search of a simple yet effective cryopreservation protocol

Cited by 60 publications

References 21 publications

Cryopreservation of testicular tissue: an alternative to maintain the reproductive capacity in different animal species

Cryopreservation of testicular tissue: an alternative to maintain the reproductive capacity in different animal species

Prepubertal testis hücre süspansiyonun dondurulmasında seeding etkisi

Optimizing methods for human testicular tissue cryopreservation and spermatogonial stem cell isolation

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