Research
Print page Print page
Switch language
Bispebjerg Hospital - a part of Copenhagen University Hospital
Published

Propagation of Spermatogonial Stem Cell-Like Cells From Infant Boys

Research output: Contribution to journalJournal articleResearchpeer-review

Harvard

Dong, L, Kristensen, SG, Hildorf, S, Gul, M, Clasen-Linde, E, Fedder, J, Hoffmann, ER, Cortes, D, Thorup, J & Andersen, CY 2019, 'Propagation of Spermatogonial Stem Cell-Like Cells From Infant Boys' Frontiers in physiology, vol. 10, 1155. https://doi.org/10.3389/fphys.2019.01155

APA

Dong, L., Kristensen, S. G., Hildorf, S., Gul, M., Clasen-Linde, E., Fedder, J., ... Andersen, C. Y. (2019). Propagation of Spermatogonial Stem Cell-Like Cells From Infant Boys. Frontiers in physiology, 10, [1155]. https://doi.org/10.3389/fphys.2019.01155

CBE

Dong L, Kristensen SG, Hildorf S, Gul M, Clasen-Linde E, Fedder J, Hoffmann ER, Cortes D, Thorup J, Andersen CY. 2019. Propagation of Spermatogonial Stem Cell-Like Cells From Infant Boys. Frontiers in physiology. 10. https://doi.org/10.3389/fphys.2019.01155

MLA

Vancouver

Dong L, Kristensen SG, Hildorf S, Gul M, Clasen-Linde E, Fedder J et al. Propagation of Spermatogonial Stem Cell-Like Cells From Infant Boys. Frontiers in physiology. 2019 Sep 19;10. 1155. https://doi.org/10.3389/fphys.2019.01155

Author

Dong, Lihua ; Kristensen, Stine Gry ; Hildorf, Simone ; Gul, Murat ; Clasen-Linde, Erik ; Fedder, Jens ; Hoffmann, Eva R ; Cortes, Dina ; Thorup, Jorgen ; Andersen, Claus Yding. / Propagation of Spermatogonial Stem Cell-Like Cells From Infant Boys. In: Frontiers in physiology. 2019 ; Vol. 10.

Bibtex

@article{1b1359dbeb8d43a18f7302bbff92f375,
title = "Propagation of Spermatogonial Stem Cell-Like Cells From Infant Boys",
abstract = "Background: Gonadotoxic treatment of malignant diseases as well as some non-malignant conditions such as cryptorchidism in young boys may result in infertility and failure to father children later in life. As a fertility preserving strategy, several centers collect testicular biopsies to cryopreserve spermatogonial stem cells (SSCs) world-wide. One of the most promising therapeutic strategies is to transplant SSCs back into the seminiferous tubules to initiate endogenous spermatogenesis. However, to obtain sufficient numbers of SSC to warrant transplantation, in vitro propagation of cells is needed together with proper validation of their stem cell identity.Materials and Methods: A minute amount of testicular biopsies (between 5 mg and 10 mg) were processed by mechanical and enzymatic digestion. SSCs were enriched by differential plating method in StemPro-34 medium supplemented with several growth factors. SSC-like cell clusters (SSCLCs) were passaged five times. SSCLCs were identified by immunohistochemical and immunofluorescence staining, using protein expression patterns in testis biopsies as reference. Quantitative polymerase chain reaction analysis of SSC markers LIN-28 homolog A (LIN28A), G antigen 1 (GAGE1), promyelocytic leukemia zinc finger protein (PLZF), integrin alpha 6 (ITGA6), ubiquitin carboxy-terminal hydrolase L1 (UCHL1) and integrin beta 1 (ITGB1) were also used to validate the SSC-like cell identity.Results: Proliferation of SSCLCs was achieved. The presence of SSCs in SSCLCs was confirmed by positive immunostaining of LIN28, UCHL1 and quantitative polymerase chain reaction for LIN28A, UCHL1, PLZF, ITGA6, and ITGB1, respectively.Conclusion: This study has demonstrated that SSCs from infant boys possess the capacity for in vitro proliferation and advance a fertility preservation strategy for pre-pubertal boys who may otherwise lose their fertility.",
author = "Lihua Dong and Kristensen, {Stine Gry} and Simone Hildorf and Murat Gul and Erik Clasen-Linde and Jens Fedder and Hoffmann, {Eva R} and Dina Cortes and Jorgen Thorup and Andersen, {Claus Yding}",
note = "Copyright {\circledC} 2019 Dong, Kristensen, Hildorf, Gul, Clasen-Linde, Fedder, Hoffmann, Cortes, Thorup and Andersen.",
year = "2019",
month = "9",
day = "19",
doi = "10.3389/fphys.2019.01155",
language = "English",
volume = "10",
journal = "Frontiers in Physiology",
issn = "1664-042X",
publisher = "Frontiers Research Foundation",

}

RIS

TY - JOUR

T1 - Propagation of Spermatogonial Stem Cell-Like Cells From Infant Boys

AU - Dong, Lihua

AU - Kristensen, Stine Gry

AU - Hildorf, Simone

AU - Gul, Murat

AU - Clasen-Linde, Erik

AU - Fedder, Jens

AU - Hoffmann, Eva R

AU - Cortes, Dina

AU - Thorup, Jorgen

AU - Andersen, Claus Yding

N1 - Copyright © 2019 Dong, Kristensen, Hildorf, Gul, Clasen-Linde, Fedder, Hoffmann, Cortes, Thorup and Andersen.

PY - 2019/9/19

Y1 - 2019/9/19

N2 - Background: Gonadotoxic treatment of malignant diseases as well as some non-malignant conditions such as cryptorchidism in young boys may result in infertility and failure to father children later in life. As a fertility preserving strategy, several centers collect testicular biopsies to cryopreserve spermatogonial stem cells (SSCs) world-wide. One of the most promising therapeutic strategies is to transplant SSCs back into the seminiferous tubules to initiate endogenous spermatogenesis. However, to obtain sufficient numbers of SSC to warrant transplantation, in vitro propagation of cells is needed together with proper validation of their stem cell identity.Materials and Methods: A minute amount of testicular biopsies (between 5 mg and 10 mg) were processed by mechanical and enzymatic digestion. SSCs were enriched by differential plating method in StemPro-34 medium supplemented with several growth factors. SSC-like cell clusters (SSCLCs) were passaged five times. SSCLCs were identified by immunohistochemical and immunofluorescence staining, using protein expression patterns in testis biopsies as reference. Quantitative polymerase chain reaction analysis of SSC markers LIN-28 homolog A (LIN28A), G antigen 1 (GAGE1), promyelocytic leukemia zinc finger protein (PLZF), integrin alpha 6 (ITGA6), ubiquitin carboxy-terminal hydrolase L1 (UCHL1) and integrin beta 1 (ITGB1) were also used to validate the SSC-like cell identity.Results: Proliferation of SSCLCs was achieved. The presence of SSCs in SSCLCs was confirmed by positive immunostaining of LIN28, UCHL1 and quantitative polymerase chain reaction for LIN28A, UCHL1, PLZF, ITGA6, and ITGB1, respectively.Conclusion: This study has demonstrated that SSCs from infant boys possess the capacity for in vitro proliferation and advance a fertility preservation strategy for pre-pubertal boys who may otherwise lose their fertility.

AB - Background: Gonadotoxic treatment of malignant diseases as well as some non-malignant conditions such as cryptorchidism in young boys may result in infertility and failure to father children later in life. As a fertility preserving strategy, several centers collect testicular biopsies to cryopreserve spermatogonial stem cells (SSCs) world-wide. One of the most promising therapeutic strategies is to transplant SSCs back into the seminiferous tubules to initiate endogenous spermatogenesis. However, to obtain sufficient numbers of SSC to warrant transplantation, in vitro propagation of cells is needed together with proper validation of their stem cell identity.Materials and Methods: A minute amount of testicular biopsies (between 5 mg and 10 mg) were processed by mechanical and enzymatic digestion. SSCs were enriched by differential plating method in StemPro-34 medium supplemented with several growth factors. SSC-like cell clusters (SSCLCs) were passaged five times. SSCLCs were identified by immunohistochemical and immunofluorescence staining, using protein expression patterns in testis biopsies as reference. Quantitative polymerase chain reaction analysis of SSC markers LIN-28 homolog A (LIN28A), G antigen 1 (GAGE1), promyelocytic leukemia zinc finger protein (PLZF), integrin alpha 6 (ITGA6), ubiquitin carboxy-terminal hydrolase L1 (UCHL1) and integrin beta 1 (ITGB1) were also used to validate the SSC-like cell identity.Results: Proliferation of SSCLCs was achieved. The presence of SSCs in SSCLCs was confirmed by positive immunostaining of LIN28, UCHL1 and quantitative polymerase chain reaction for LIN28A, UCHL1, PLZF, ITGA6, and ITGB1, respectively.Conclusion: This study has demonstrated that SSCs from infant boys possess the capacity for in vitro proliferation and advance a fertility preservation strategy for pre-pubertal boys who may otherwise lose their fertility.

U2 - 10.3389/fphys.2019.01155

DO - 10.3389/fphys.2019.01155

M3 - Journal article

VL - 10

JO - Frontiers in Physiology

JF - Frontiers in Physiology

SN - 1664-042X

M1 - 1155

ER -

ID: 58140104