Short-term hypothermic preservation of human testicular tissue: the effect of storage medium and storage period

Faes, Katrien; Goossens, Ellen

doi:10.1016/j.fertnstert.2016.01.018

Cited by 13 publications

(12 citation statements)

References 27 publications

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“…A previous study reported that immature non-human primate testicular tissues that had been cold stored for 24 hours could be xenografted and initiate spermatogenesis up to the spermatocyte stage (Jahnukainen et al ., 2007). Human testicular tissues have been cold stored for up to 3 days at 4°C without altering tissue morphology, Sertoli cell morphology, number of spermatogonia, or number of apoptotic cells (Faes and Goossens, 2016). Collectively, these data suggest that some period of cold storage during shipping may be acceptable.…”

Section: Discussionmentioning

confidence: 99%

Testicular tissue cryopreservation: 8 years of experience from a coordinated network of academic centers

et al. 2019

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STUDY QUESTION Is it feasible to disseminate testicular tissue cryopreservation with a standardized protocol through a coordinated network of centers and provide centralized processing/freezing for centers that do not have those capabilities? SUMMARY ANSWER Centralized processing and freezing of testicular tissue from multiple sites is feasible and accelerates recruitment, providing the statistical power to make inferences that may inform fertility preservation practice. WHAT IS KNOWN ALREADY Several centers in the USA and abroad are preserving testicular biopsies for patients who cannot preserve sperm in anticipation that cell- or tissue-based therapies can be used in the future to generate sperm and offspring. STUDY DESIGN, SIZE, DURATION Testicular tissue samples from 189 patients were cryopreserved between January 2011 and November 2018. Medical diagnosis, previous chemotherapy exposure, tissue weight, and presence of germ cells were recorded. PARTICIPANTS/MATERIALS, SETTING, METHODS Human testicular tissue samples were obtained from patients undergoing treatments likely to cause infertility. Twenty five percent of the patient’s tissue was donated to research and 75% was stored for patient’s future use. The tissue was weighed, and research tissue was fixed for histological analysis with Periodic acid-Schiff hematoxylin staining and/or immunofluorescence staining for DEAD-box helicase 4, and/or undifferentiated embryonic cell transcription factor 1. MAIN RESULTS AND THE ROLE OF CHANCE The average age of fertility preservation patients was 7.9 (SD = 5) years and ranged from 5 months to 34 years. The average amount of tissue collected was 411.3 (SD = 837.3) mg and ranged from 14.4 mg—6880.2 mg. Malignancies ( n = 118) were the most common indication for testicular tissue freezing, followed by blood disorders ( n = 45) and other conditions ( n = 26). Thirty nine percent ( n = 74) of patients had initiated their chemotherapy prior to undergoing testicular biopsy. Of the 189 patients recruited to date, 137 have been analyzed for the presence of germ cells and germ cells were confirmed in 132. LIMITATIONS, REASONS FOR CAUTION This is a descriptive study of testicular tissues obtained from patients who were at risk of infertility. The function of spermatogonia in those biopsies could not be tested by transplantation due limited sample size. WIDER IMPLICATIONS OF THE FINDINGS Patients and/or guardians are willing to pursue an experimental fertility preservation procedure when no alternatives are available. Our coordinated network of centers found that many patients request fertility preservation after initiating gonadotoxic therapies. This study demonstrates that undifferentiated...

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Section: Discussionmentioning

confidence: 99%

Testicular tissue cryopreservation: 8 years of experience from a coordinated network of academic centers

et al. 2019

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“…Thus, testicular cryopreservation is a reproductive biotechnology of great importance in the recovery and conservation of genetic material from prepubertal animals belonging to endangered wild species (Pukazhenthi et al., ), as well as in the preservation of reproductive potential of animals with higher zootechnical importance, which have died early or who have undergone treatments that cause infertility, or for patients suffering from testicular tumours undergoing chemotherapy or radiotherapy (Faes & Goossens, ).…”

Section: Introductionmentioning

confidence: 99%

Different associations of cryoprotectants for testicular tissue of prepubertal cats submitted to vitrification

Lima

Silva

Morais

et al. 2016

Reprod Domestic Animals

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The cryopreservation of testicular tissue is presented as the only alternative for the preservation of genetic material from prepubertal animals. However, this biotechnology is still being tested. The objective of this study was to evaluate the effect of different associations of cryoprotectants and the potential of cell proliferation after vitrification of testicular tissue of prepubertal cats. Five testicular pairs from five prepubertal cats were used, and each pair was divided into four fragments. Of these, one fragment composed of the control group (CG) and the rest were distributed in experimental groups according to the associations of cryoprotectants to be tested (dimethyl sulphoxide (DMSO)/glycerol (GLY); ethylene glycol (EG)/GLY) or DMSO/EG) in a final cryoprotectant concentration of 5.6 m. The fragments were submitted to vitrification, and after one week, fragments were heated and processed for histomorphological evaluation and quantification of nucleolar organizer regions (NORs). DMSO/GLY did not differ from CG and was superior to the other vitrified groups, as to cell separation and degree of shrinkage of the basal membrane. Concerning cell differentiation, visibility of the nucleus and nuclear condensation, all the vitrified groups were inferior to CG; however, DMSO/EG was inferior to DMSO/GLY and EG/GLY, which did not differ among themselves. CG was superior to all groups in quantification of NORs. DMSO/EG was inferior to all others, and there was no difference between DMSO/GLY and EG/GLY. The association DMSO/GLY presented the best preservation of tissue integrity and potential of cell proliferation after vitrification of the testicular tissue of prepubertal cats.

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“…This raises concern about the quality of ITT when shipment to the banking facility is delayed. Earlier study in human ITT showed no change in the structural integrity up to 3 days of holding at 4 °C [ 13 ]. However, in our study, after 14 days of in vitro culture, the cell viability, testosterone production, and the number of postmeiotic population were identical across both ultraprofound and profound-hypothermic-temperature groups and at both 6 h holding, but significantly reduced in 24 h holding at profound-hypothermic-temperature.…”

Section: Discussionmentioning

confidence: 99%

“…If the retrieved ITT can be cryopreserved with minimal manipulation, the chance to recover optimum number of cells for fertility restoration techniques might increase [10]. Hence, earlier studies have addressed the effects of varying tissue size, storage temperatures, and storage periods in porcine model [10] and in human [11][12][13][14], to recommend the ideal conditions. No loss of cell viability and structural integrity was observed during the length of cooling [10], and proliferative potential was unaltered when cooled tissue was thawed and xenografted [15].…”

Section: Introductionmentioning

confidence: 99%

“…No loss of cell viability and structural integrity was observed during the length of cooling [ 10 ], and proliferative potential was unaltered when cooled tissue was thawed and xenografted [ 15 ]. On the other hand, human ITT preservation at 4 °C for 3 days did not impair tissue integrity, Sertoli cell morphology, stem cell population, and the incidence of apoptosis [ 13 ]. In vitro maturation of human germ cells from ITT is being attempted as a promising alternative to in vivo approaches [ 11 , 16 – 19 ], hence handling and manipulation of ITT may play an important role in maintaining the functionality of cells.…”

Section: Introductionmentioning

confidence: 99%

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Impact of Temperature and Time Interval Prior to Immature Testicular-Tissue Organotypic Culture on Cellular Niche

et al. 2020

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Cryopreservation of immature-testicular-tissue (ITT) prior to gonadotoxic treatment, while experimental, is the only recommended option for fertility preservation in prepubertal boys. The handling and manipulation of ITT before cryopreservation could influence the functionality of cells during fertility restoration, which this study explored by evaluating cellular niche and quality of mouse ITT subjected to various temperatures and time durations in vitro. ITT from 6-day-old mice were handled at ultraprofound-hypothermic, profound-hypothermic, and mild-warm-ischemic temperatures for varying time periods prior to 14-day organotypic culture. Viability, functionality, synaptonemal complex and chromatin remodeling markers were assessed. Results have shown that cell viability, testosterone level, and in vitro proliferation ability did not change when ITT were held at ultraprofound-hypothermic-temperature up to 24 h, whereas cell viability was significantly reduced (P < 0.01), when held at profound-hypothermic-temperature for 24 h before culture. Further, cell viability and testosterone levels in cultured cells from profound-hypothermic group were comparable to corresponding ultraprofound-hypothermic group but with moderate reduction in postmeiotic cells (P < 0.01). In conclusion, holding ITT at ultraprofound-hypothermic-temperature is most suitable for organotypic culture, whereas short-term exposure at profound-hypothermic-temperature may compromise postmeiotic germ cell yield post in vitro culture. This data, albeit in mouse model, will have immense value in human prepubertal fertility restoration research.

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Short-term hypothermic preservation of human testicular tissue: the effect of storage medium and storage period

Cited by 13 publications

References 27 publications

Testicular tissue cryopreservation: 8 years of experience from a coordinated network of academic centers

Testicular tissue cryopreservation: 8 years of experience from a coordinated network of academic centers

Different associations of cryoprotectants for testicular tissue of prepubertal cats submitted to vitrification

Impact of Temperature and Time Interval Prior to Immature Testicular-Tissue Organotypic Culture on Cellular Niche

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