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The effects of poly L-lactic acid nanofiber scaffold on mouse spermatogonial stem cell culture

INTRODUCTION: A 3D-nanofiber scaffold acts in a similar way to the extracellular matrix (ECM)/basement membrane that enhances the proliferation and self-renewal of stem cells. The goal of the present study was to investigate the effects of a poly L-lactic acid (PLLA) nanofiber scaffold on frozen-tha...

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Autores principales: Eslahi, Neda, Hadjighassem, Mahmoud Reza, Joghataei, Mohammad Taghi, Mirzapour, Tooba, Bakhtiyari, Mehrdad, Shakeri, Malak, Pirhajati, Vahid, Shirinbayan, Peymaneh, Koruji, Morteza
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Dove Medical Press 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3848747/
https://www.ncbi.nlm.nih.gov/pubmed/24348035
http://dx.doi.org/10.2147/IJN.S45535
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author Eslahi, Neda
Hadjighassem, Mahmoud Reza
Joghataei, Mohammad Taghi
Mirzapour, Tooba
Bakhtiyari, Mehrdad
Shakeri, Malak
Pirhajati, Vahid
Shirinbayan, Peymaneh
Koruji, Morteza
author_facet Eslahi, Neda
Hadjighassem, Mahmoud Reza
Joghataei, Mohammad Taghi
Mirzapour, Tooba
Bakhtiyari, Mehrdad
Shakeri, Malak
Pirhajati, Vahid
Shirinbayan, Peymaneh
Koruji, Morteza
author_sort Eslahi, Neda
collection PubMed
description INTRODUCTION: A 3D-nanofiber scaffold acts in a similar way to the extracellular matrix (ECM)/basement membrane that enhances the proliferation and self-renewal of stem cells. The goal of the present study was to investigate the effects of a poly L-lactic acid (PLLA) nanofiber scaffold on frozen-thawed neonate mouse spermatogonial stem cells (SSCs) and testis tissues. METHODS: The isolated spermatogonial cells were divided into six culture groups: (1) fresh spermatogonial cells, (2) fresh spermatogonial cells seeded onto PLLA, (3) frozen-thawed spermatogonial cells, (4) frozen-thawed spermatogonial cells seeded onto PLLA, (5) spermatogonial cells obtained from frozen-thawed testis tissue, and (6) spermatogonial cells obtained from frozen-thawed testis tissue seeded onto PLLA. Spermatogonial cells and testis fragments were cryopreserved and cultured for 3 weeks. Cluster assay was performed during the culture. The presence of spermatogonial cells in the culture was determined by a reverse transcriptase polymerase chain reaction for spermatogonial markers (Oct4, GFRα-1, PLZF, Mvh(VASA), Itgα6, and Itgβ1), as well as the ultrastructural study of cell clusters and SSCs transplantation to a recipient azoospermic mouse. The significance of the data was analyzed using the repeated measures and analysis of variance. RESULTS: The findings indicated that the spermatogonial cells seeded on PLLA significantly increased in vitro spermatogonial cell cluster formations in comparison with the control groups (culture of SSCs not seeded on PLLA) (P≤0.001). The viability rate for the frozen cells after thawing was 63.00% ± 3.56%. This number decreased significantly (40.00% ± 0.82%) in spermatogonial cells obtained from the frozen-thawed testis tissue. Both groups, however, showed in vitro cluster formation. Although the expression of spermatogonial markers was maintained after 3 weeks of culture, there was a significant downregulation for some spermatogonial genes in the experimental groups compared with those of the control groups. Furthermore, transplantation assay and transmission electron microscopy studies suggested the presence of SSCs among the cultured cells. CONCLUSION: Although PLLA can increase the in vitro cluster formation of neonate fresh and frozen-thawed spermatogonial cells, it may also cause them to differentiate during cultivation. The study therefore has implications for SSCs proliferation and germ cell differentiation in vitro.
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spelling pubmed-38487472013-12-13 The effects of poly L-lactic acid nanofiber scaffold on mouse spermatogonial stem cell culture Eslahi, Neda Hadjighassem, Mahmoud Reza Joghataei, Mohammad Taghi Mirzapour, Tooba Bakhtiyari, Mehrdad Shakeri, Malak Pirhajati, Vahid Shirinbayan, Peymaneh Koruji, Morteza Int J Nanomedicine Original Research INTRODUCTION: A 3D-nanofiber scaffold acts in a similar way to the extracellular matrix (ECM)/basement membrane that enhances the proliferation and self-renewal of stem cells. The goal of the present study was to investigate the effects of a poly L-lactic acid (PLLA) nanofiber scaffold on frozen-thawed neonate mouse spermatogonial stem cells (SSCs) and testis tissues. METHODS: The isolated spermatogonial cells were divided into six culture groups: (1) fresh spermatogonial cells, (2) fresh spermatogonial cells seeded onto PLLA, (3) frozen-thawed spermatogonial cells, (4) frozen-thawed spermatogonial cells seeded onto PLLA, (5) spermatogonial cells obtained from frozen-thawed testis tissue, and (6) spermatogonial cells obtained from frozen-thawed testis tissue seeded onto PLLA. Spermatogonial cells and testis fragments were cryopreserved and cultured for 3 weeks. Cluster assay was performed during the culture. The presence of spermatogonial cells in the culture was determined by a reverse transcriptase polymerase chain reaction for spermatogonial markers (Oct4, GFRα-1, PLZF, Mvh(VASA), Itgα6, and Itgβ1), as well as the ultrastructural study of cell clusters and SSCs transplantation to a recipient azoospermic mouse. The significance of the data was analyzed using the repeated measures and analysis of variance. RESULTS: The findings indicated that the spermatogonial cells seeded on PLLA significantly increased in vitro spermatogonial cell cluster formations in comparison with the control groups (culture of SSCs not seeded on PLLA) (P≤0.001). The viability rate for the frozen cells after thawing was 63.00% ± 3.56%. This number decreased significantly (40.00% ± 0.82%) in spermatogonial cells obtained from the frozen-thawed testis tissue. Both groups, however, showed in vitro cluster formation. Although the expression of spermatogonial markers was maintained after 3 weeks of culture, there was a significant downregulation for some spermatogonial genes in the experimental groups compared with those of the control groups. Furthermore, transplantation assay and transmission electron microscopy studies suggested the presence of SSCs among the cultured cells. CONCLUSION: Although PLLA can increase the in vitro cluster formation of neonate fresh and frozen-thawed spermatogonial cells, it may also cause them to differentiate during cultivation. The study therefore has implications for SSCs proliferation and germ cell differentiation in vitro. Dove Medical Press 2013 2013-11-27 /pmc/articles/PMC3848747/ /pubmed/24348035 http://dx.doi.org/10.2147/IJN.S45535 Text en © 2013 Eslahi et al. This work is published by Dove Medical Press Limited, and licensed under Creative Commons Attribution – Non Commercial (unported, v3.0) License The full terms of the License are available at http://creativecommons.org/licenses/by-nc/3.0/. Non-commercial uses of the work are permitted without any further permission from Dove Medical Press Limited, provided the work is properly attributed.
spellingShingle Original Research
Eslahi, Neda
Hadjighassem, Mahmoud Reza
Joghataei, Mohammad Taghi
Mirzapour, Tooba
Bakhtiyari, Mehrdad
Shakeri, Malak
Pirhajati, Vahid
Shirinbayan, Peymaneh
Koruji, Morteza
The effects of poly L-lactic acid nanofiber scaffold on mouse spermatogonial stem cell culture
title The effects of poly L-lactic acid nanofiber scaffold on mouse spermatogonial stem cell culture
title_full The effects of poly L-lactic acid nanofiber scaffold on mouse spermatogonial stem cell culture
title_fullStr The effects of poly L-lactic acid nanofiber scaffold on mouse spermatogonial stem cell culture
title_full_unstemmed The effects of poly L-lactic acid nanofiber scaffold on mouse spermatogonial stem cell culture
title_short The effects of poly L-lactic acid nanofiber scaffold on mouse spermatogonial stem cell culture
title_sort effects of poly l-lactic acid nanofiber scaffold on mouse spermatogonial stem cell culture
topic Original Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3848747/
https://www.ncbi.nlm.nih.gov/pubmed/24348035
http://dx.doi.org/10.2147/IJN.S45535
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