Testicular tissue cryopreservation: 8 years of experience from a coordinated network of academic centers
Hanna Valli-Pulaski,Karen A. Peters,Kathrin Gassei,Sarah Steimer,Meena Sukhwani,Brian P. Hermann,L Dwomor,Sherin David,Adetunji P. Fayomi,Sarah Munyoki,Tianjiao Chu,R Chaudhry,Glenn M. Cannon,P J Fox,Thomas M. Jaffe,Joseph S. Sanfilippo,Marie N. Menke,Eitan Lunenfeld,Maram Abofoul-Azab,Leonard S. Sender,J Messina,L M Klimpel,Yasmin Gosiengfiao,Erin E. Rowell,Michael H. Hsieh,Candace F. Granberg,Pramod P. Reddy,Jay I. Sandlow,Mahmoud Huleihel,Kyle E. Orwig +29 more
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TLDR
This study demonstrates that undifferentiated stem and progenitor spermatogonia may be recovered from the testicular tissues of patients who are in the early stages of their treatment and have not yet received an ablative dose of therapy.Abstract:
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 stem and progenitor spermatogonia may be recovered from the testicular tissues of patients who are in the early stages of their treatment and have not yet received an ablative dose of therapy. The function of those spermatogonia was not tested. Study funding/competing interest(s) Support for the research was from the Eunice Kennedy Shriver National Institute for Child Health and Human Development grants HD061289 and HD092084, the Scaife Foundation, the Richard King Mellon Foundation, the Departments of Ob/Gyn & Reproductive Sciences and Urology of the University of Pittsburgh Medical Center, United States-Israel Binational Science Foundation (BSF), and the Kahn Foundation. The authors declare that they do not have competing financial interests.read more
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Fertility preservation in boys: recent developments and new insights †
Ellen Goossens,Kirsi Jahnukainen,Kirsi Jahnukainen,Rod T. Mitchell,Amm van Pelt,Guido Pennings,N Rives,J Poels,Christine Wyns,Sheila Lane,Kenny A. Rodriguez-Wallberg,Kenny A. Rodriguez-Wallberg,A Rives,Hanna Valli-Pulaski,Sarah Steimer,Sabine Kliesch,Aude Braye,María del Mar Andrés,J Medrano,Liliana Ramos,SG Kristensen,Claus Yding Andersen,Ragnar Bjarnason,Kyle E. Orwig,Nina Neuhaus,Jan-Bernd Stukenborg +25 more
TL;DR: This review aims to provide an update on clinical practices and experimental methods, as well as to describe patient management inclusion strategies used to preserve and restore the fertility of prepubertal boys at high risk of fertility loss.
Journal ArticleDOI
Tissue Engineering to Improve Immature Testicular Tissue and Cell Transplantation Outcomes: One Step Closer to Fertility Restoration for Prepubertal Boys Exposed to Gonadotoxic Treatments.
Federico Del Vento,Maxime Vermeulen,Francesca de Michele,Francesca de Michele,Maria Grazia Giudice,Maria Grazia Giudice,Jonathan Poels,Anne des Rieux,Christine Wyns,Christine Wyns +9 more
TL;DR: The benefits that might be gathered using bioengineering techniques to enhance transplantation outcomes by optimizing early tissue graft revascularization, protecting cells from toxic insults linked to ischemic injury and exploring strategies to promote cellular differentiation are focused on.
Journal ArticleDOI
Generation of Organized Porcine Testicular Organoids in Solubilized Hydrogels from Decellularized Extracellular Matrix.
Maxime Vermeulen,Federico Del Vento,Marc Kanbar,Sébastien Pyr dit Ruys,Didier Vertommen,Jonathan Poels,Christine Wyns,Christine Wyns +7 more
TL;DR: Generation of testicular organoids (TOs) after cell selection is a novel strategy aimed at restoring fertility in prepubertal boys about to undergo cancer therapies and suggests a better preservation of growth factors within TOs developed from decellularized ITT and thus a better potential to restore the reproductive capacity.
Journal ArticleDOI
Fertility preservation for prepubertal boys: lessons learned from the past and update on remaining challenges towards clinical translation
Christine Wyns,Christine Wyns,Marc Kanbar,Marc Kanbar,Maria Grazia Giudice,Maria Grazia Giudice,Jonathan Poels,Jonathan Poels +7 more
TL;DR: Preliminary follow-up data indicate that around 27% of boys who have undergone testicular sampling as an FP measure have proved azoospermic and must therefore solely rely on their cryostored ITT to ensure biologic parenthood.
Journal ArticleDOI
Review of injection techniques for spermatogonial stem cell transplantation.
Murat Gul,Murat Gul,Simone Hildorf,Lihua Dong,Jorgen Thorup,Eva Hoffmann,Christian Fuglesang S Jensen,Jens Sønksen,Dina Cortes,Dina Cortes,Jens Fedder,Jens Fedder,Claus Yding Andersen,Claus Yding Andersen,Ellen Goossens +14 more
TL;DR: This narrative review provides an overview of the currently used experimental injection techniques for SSC transplantation in animal and human testes to provide insight into the mechanisms of successful assisted human reproduction.
References
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Journal ArticleDOI
Fertility Preservation in Patients With Cancer: ASCO Clinical Practice Guideline Update.
Kutluk Oktay,Brittany E. Harvey,Ann H. Partridge,Gwendolyn P. Quinn,Joyce Reinecke,Hugh S. Taylor,W. Hamish B. Wallace,Erica T. Wang,Alison W. Loren +8 more
TL;DR: There is conflicting evidence to recommend gonadotrophin-releasing hormone agonists (GnRHa) and other means of ovarian suppression for fertility preservation and the panel notes that the field of ovarian tissue cryopreservation is advancing quickly and may evolve to become standard therapy in the future.
Journal ArticleDOI
Cancer and fertility preservation: international recommendations from an expert meeting
Matteo Lambertini,Lucia Del Mastro,Maria Carolina Pescio,Claus Yding Andersen,Hatem A. Azim,Fedro A. Peccatori,Mauro Costa,Alberto Revelli,Francesca Salvagno,Alessandra Gennari,Filippo Ubaldi,Giovanni Battista La Sala,Cristofaro De Stefano,Hamish Wallace,Ann H. Partridge,Paola Anserini +15 more
TL;DR: Although there is a great interest in this field, due to the lack of large prospective cohort studies and randomized trials on these topics, the level of evidence is not higher than 3 for most of the recommendations highlighting the need of further research efforts in many areas of this field.
Journal ArticleDOI
Propagation of human spermatogonial stem cells in vitro.
Hooman Sadri-Ardekani,S.C. Mizrak,Saskia K.M. van Daalen,Cindy M. Korver,Hermien L. Roepers-Gajadien,Morteza Koruji,Suzanne Hovingh,Theo M. de Reijke,Jean J.M.C.H. de la Rosette,Fulco van der Veen,Dirk G. de Rooij,Sjoerd Repping,Ans M.M. van Pelt +12 more
Abstract: Context Young boys treated with high-dose chemotherapy are often confronted with infertility once they reach adulthood. Cryopreserving testicular tissue before chemotherapy and autotransplantation of spermatogonial stem cells at a later stage could theoretically allow for restoration of fertility. Objective To establish in vitro propagation of human spermatogonial stem cells from small testicular biopsies to obtain an adequate number of cells for successful transplantation. Design, Setting, and Participants Study performed from April 2007 to July 2009 using testis material donated by 6 adult men who underwent orchidectomy as part of prostate cancer treatment. Testicular cells were isolated and cultured in supplemented StemPro medium; germline stem cell clusters that arose were subcultured on human placental laminin–coated dishes in the same medium. Presence of spermatogonia was determined by reverse transcriptase polymerase chain reaction and immunofluorescence for spermatogonial markers. To test for the presence of functional spermatogonial stem cells in culture, xenotransplantation to testes of immunodeficient mice was performed, and migrated human spermatogonial stem cells after transplantation were detected by COT-1 fluorescence in situ hybridization. The number of colonized spermatogonial stem cells transplanted at early and later points during culture were counted to determine propagation. Main Outcome Measures Propagation of spermatogonial stem cells over time. Results Testicular cells could be cultured and propagated up to 15 weeks. Germline stem cell clusters arose in the testicular cell cultures from all 6 men and could be subcultured and propagated up to 28 weeks. Expression of spermatogonial markers on both the RNA and protein level was maintained throughout the entire culture period. In 4 of 6 men, xenotransplantation to mice demonstrated the presence of functional spermatogonial stem cells, even after prolonged in vitro culture. Spermatogonial stem cell numbers increased 53-fold within 19 days in the testicular cell culture and increased 18 450-fold within 64 days in the germline stem cell subculture. Conclusion Long-term culture and propagation of human spermatogonial stem cells in vitro is achievable.
Journal ArticleDOI
Fertility of Male Survivors of Childhood Cancer: A Report From the Childhood Cancer Survivor Study
Daniel M. Green,Toana Kawashima,Marilyn Stovall,Wendy M. Leisenring,Charles A. Sklar,Ann C. Mertens,Sarah S. Donaldson,Julianne Byrne,Leslie L. Robison +8 more
TL;DR: This large study identified risk factors for decreased fertility that may be used for counseling male cancer patients.
Journal ArticleDOI
The Cyclophosphamide Equivalent Dose as an Approach for Quantifying Alkylating Agent Exposure. A Report from the Childhood Cancer Survivor Study
Daniel M. Green,Vikki G. Nolan,Pamela Goodman,John Whitton,Deo Kumar Srivastava,Wendy M. Leisenring,Joseph P. Neglia,Charles A. Sklar,Sue C. Kaste,Melissa M. Hudson,Lisa Diller,Marilyn Stovall,Sarah S. Donaldson,Leslie L. Robison +13 more
TL;DR: The widely used Alkylating Agent Dose (AAD) score is derived from the drug dose distribution of the study population and thus cannot be used for comparisons across populations as each will have a unique distribution of drug doses.
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