In Vitro Propagation of XXY Undifferentiated Mouse Spermatogonia: Model for Fertility Preservation in Klinefelter Syndrome Patients
Klinefelter syndrome (KS) is characterized by a masculine phenotype, supernumerary sex chromosomes (usually XXY), and spermatogonial stem cell (SSC) loss in their early life. Affecting 1 out of every 650 males born, KS is the most common genetic cause of male infertility, and new fertility preservat...
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Published in | International journal of molecular sciences Vol. 23; no. 1; p. 173 |
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Main Authors | , , , , , , , , , |
Format | Journal Article |
Language | English |
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24.12.2021
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Abstract | Klinefelter syndrome (KS) is characterized by a masculine phenotype, supernumerary sex chromosomes (usually XXY), and spermatogonial stem cell (SSC) loss in their early life. Affecting 1 out of every 650 males born, KS is the most common genetic cause of male infertility, and new fertility preservation strategies are critically important for these patients. In this study, testes from 41, XXY prepubertal (3-day-old) mice were frozen-thawed. Isolated testicular cells were cultured and characterized by qPCR, digital PCR, and flow cytometry analyses. We demonstrated that SSCs survived and were able to be propagated with testicular somatic cells in culture for up to 120 days. DNA fluorescent in situ hybridization (FISH) showed the presence of XXY spermatogonia at the beginning of the culture and a variety of propagated XY, XX, and XXY spermatogonia at the end of the culture. These data provide the first evidence that an extra sex chromosome was lost during innate SSC culture, a crucial finding in treating KS patients for preserving and propagating SSCs for future sperm production, either in vitro or in vivo. This in vitro propagation system can be translated to clinical fertility preservation for KS patients. |
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AbstractList | Klinefelter syndrome (KS) is characterized by a masculine phenotype, supernumerary sex chromosomes (usually XXY), and spermatogonial stem cell (SSC) loss in their early life. Affecting 1 out of every 650 males born, KS is the most common genetic cause of male infertility, and new fertility preservation strategies are critically important for these patients. In this study, testes from 41, XXY prepubertal (3-day-old) mice were frozen-thawed. Isolated testicular cells were cultured and characterized by qPCR, digital PCR, and flow cytometry analyses. We demonstrated that SSCs survived and were able to be propagated with testicular somatic cells in culture for up to 120 days. DNA fluorescent in situ hybridization (FISH) showed the presence of XXY spermatogonia at the beginning of the culture and a variety of propagated XY, XX, and XXY spermatogonia at the end of the culture. These data provide the first evidence that an extra sex chromosome was lost during innate SSC culture, a crucial finding in treating KS patients for preserving and propagating SSCs for future sperm production, either in vitro or in vivo. This in vitro propagation system can be translated to clinical fertility preservation for KS patients. |
Author | Zarandi, Nima Pourhabibi Swerdloff, Ronald S Pettenati, Mark J Galdon, Guillermo Deebel, Nicholas A Sadri-Ardekani, Hooman Atala, Anthony Wang, Christina Kogan, Stanley Lue, Yanhe |
AuthorAffiliation | 2 Facultad de Medicina Universidad de Barcelona, 08036 Barcelona, Spain 3 Department of Urology, Wake Forest School of Medicine, Winston-Salem, NC 27101, USA 1 Wake Forest Institute for Regenerative Medicine (WFIRM), Wake Forest School of Medicine, Winston-Salem, NC 27101, USA; galdon.guillermo@gmail.com (G.G.); ndeebel@wakehealth.edu (N.A.D.); npourhab@wakehealth.edu (N.P.Z.); skogan@wakehealth.edu (S.K.); aatala@wakehealth.edu (A.A.) 4 Section of Medical Genetics, Department of Pathology, Wake Forest School of Medicine, Winston-Salem, NC 27101, USA; pettenat@wakehealth.edu 5 Division of Endocrinology, The Lundquist Institute and Harbor-UCLA Medical Center, Torrance, CA 90502, USA; wang@lundquist.org (C.W.); swerdloff@lundquist.org (R.S.S.); Lue@lundquist.org (Y.L.) |
AuthorAffiliation_xml | – name: 2 Facultad de Medicina Universidad de Barcelona, 08036 Barcelona, Spain – name: 4 Section of Medical Genetics, Department of Pathology, Wake Forest School of Medicine, Winston-Salem, NC 27101, USA; pettenat@wakehealth.edu – name: 5 Division of Endocrinology, The Lundquist Institute and Harbor-UCLA Medical Center, Torrance, CA 90502, USA; wang@lundquist.org (C.W.); swerdloff@lundquist.org (R.S.S.); Lue@lundquist.org (Y.L.) – name: 3 Department of Urology, Wake Forest School of Medicine, Winston-Salem, NC 27101, USA – name: 1 Wake Forest Institute for Regenerative Medicine (WFIRM), Wake Forest School of Medicine, Winston-Salem, NC 27101, USA; galdon.guillermo@gmail.com (G.G.); ndeebel@wakehealth.edu (N.A.D.); npourhab@wakehealth.edu (N.P.Z.); skogan@wakehealth.edu (S.K.); aatala@wakehealth.edu (A.A.) |
Author_xml | – sequence: 1 givenname: Guillermo orcidid: 0000-0003-4409-7652 surname: Galdon fullname: Galdon, Guillermo organization: Facultad de Medicina Universidad de Barcelona, 08036 Barcelona, Spain – sequence: 2 givenname: Nicholas A surname: Deebel fullname: Deebel, Nicholas A organization: Department of Urology, Wake Forest School of Medicine, Winston-Salem, NC 27101, USA – sequence: 3 givenname: Nima Pourhabibi surname: Zarandi fullname: Zarandi, Nima Pourhabibi organization: Wake Forest Institute for Regenerative Medicine (WFIRM), Wake Forest School of Medicine, Winston-Salem, NC 27101, USA – sequence: 4 givenname: Mark J surname: Pettenati fullname: Pettenati, Mark J organization: Section of Medical Genetics, Department of Pathology, Wake Forest School of Medicine, Winston-Salem, NC 27101, USA – sequence: 5 givenname: Stanley surname: Kogan fullname: Kogan, Stanley organization: Department of Urology, Wake Forest School of Medicine, Winston-Salem, NC 27101, USA – sequence: 6 givenname: Christina surname: Wang fullname: Wang, Christina organization: Division of Endocrinology, The Lundquist Institute and Harbor-UCLA Medical Center, Torrance, CA 90502, USA – sequence: 7 givenname: Ronald S surname: Swerdloff fullname: Swerdloff, Ronald S organization: Division of Endocrinology, The Lundquist Institute and Harbor-UCLA Medical Center, Torrance, CA 90502, USA – sequence: 8 givenname: Anthony orcidid: 0000-0001-8186-2160 surname: Atala fullname: Atala, Anthony organization: Department of Urology, Wake Forest School of Medicine, Winston-Salem, NC 27101, USA – sequence: 9 givenname: Yanhe surname: Lue fullname: Lue, Yanhe organization: Division of Endocrinology, The Lundquist Institute and Harbor-UCLA Medical Center, Torrance, CA 90502, USA – sequence: 10 givenname: Hooman orcidid: 0000-0003-4377-4404 surname: Sadri-Ardekani fullname: Sadri-Ardekani, Hooman organization: Department of Urology, Wake Forest School of Medicine, Winston-Salem, NC 27101, USA |
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CitedBy_id | crossref_primary_10_3389_fendo_2022_1002279 crossref_primary_10_1016_j_fertnstert_2022_07_015 crossref_primary_10_1016_j_euf_2022_11_007 crossref_primary_10_1136_archdischild_2020_320831 |
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Keywords | Klinefelter syndrome spermatogonia fertility preservation spermatogonia stem cells male infertility |
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Snippet | Klinefelter syndrome (KS) is characterized by a masculine phenotype, supernumerary sex chromosomes (usually XXY), and spermatogonial stem cell (SSC) loss in... |
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SubjectTerms | Animals Biopsy Cell culture Chromosomes Cryopreservation Disease Models, Animal Fertility Fertility Preservation Flow cytometry Fluorescence Fluorescence in situ hybridization Gene expression Hyperplasia In vitro fertilization Infertility Klinefelter Syndrome Klinefelter's syndrome Male male infertility Mice Patients Phenotypes Population Preservation Propagation Puberty Semen Preservation Sex chromosomes Somatic cells Sperm Spermatogonia spermatogonia stem cells Stem cells Supernumerary Testes Testosterone |
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Title | In Vitro Propagation of XXY Undifferentiated Mouse Spermatogonia: Model for Fertility Preservation in Klinefelter Syndrome Patients |
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