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Separating mouse malignant cell line (EL4) from neonate spermatogonial stem cells utilizing microfluidic device in vitro

BACKGROUND: Some children who have survived cancer will be azoospermic in the future. Performing isolation and purification procedures for spermatogonial stem cells (SSC) is very critical. In this regard, performing the process of decontamination of cancerous cells is the initial step. The major obj...

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Autores principales: Ashtari, Behnaz, Shams, Azar, Esmaeilzadeh, Narges, Tanbakooei, Sara, Koruji, Morteza, Moghadam, Mojtaba Johari, Ansari, Javad Mohajer, Moghadam, Adel Johari, Shabani, Ronak
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7245899/
https://www.ncbi.nlm.nih.gov/pubmed/32448280
http://dx.doi.org/10.1186/s13287-020-01671-1
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author Ashtari, Behnaz
Shams, Azar
Esmaeilzadeh, Narges
Tanbakooei, Sara
Koruji, Morteza
Moghadam, Mojtaba Johari
Ansari, Javad Mohajer
Moghadam, Adel Johari
Shabani, Ronak
author_facet Ashtari, Behnaz
Shams, Azar
Esmaeilzadeh, Narges
Tanbakooei, Sara
Koruji, Morteza
Moghadam, Mojtaba Johari
Ansari, Javad Mohajer
Moghadam, Adel Johari
Shabani, Ronak
author_sort Ashtari, Behnaz
collection PubMed
description BACKGROUND: Some children who have survived cancer will be azoospermic in the future. Performing isolation and purification procedures for spermatogonial stem cells (SSC) is very critical. In this regard, performing the process of decontamination of cancerous cells is the initial step. The major objective of the present study is to separate the malignant EL4 cell line in mice and spermatogonial stem cells in vitro. METHODS: The spermatogonial stem cells of sixty neonatal mice were isolated, and the procedure of co-culturing was carried out by EL4 which were classified into 2 major groups: (1) the control group (co-culture in a growth medium) and (2) the group of co-cultured cells which were separated using the microfluidic device. The percentage of cells was assessed using flow cytometry technique and common laboratory technique of immunocytochemistry and finally was confirmed through the laboratory technique of reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: The actual percentage of EL4 and SSC after isolation was collected at two outlets: the outputs for the smaller outlet were 0.12% for SSC and 42.14% for EL4, while in the larger outlet, the outputs were 80.38% for SSC and 0.32% for EL4; in the control group, the percentages of cells were 21.44% for SSC and 23.28% for EL4 (based on t test (p ≤ 0.05)). CONCLUSIONS: The present study demonstrates that the use of the microfluidic device is effective in separating cancer cells from spermatogonial stem cells.
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spelling pubmed-72458992020-06-01 Separating mouse malignant cell line (EL4) from neonate spermatogonial stem cells utilizing microfluidic device in vitro Ashtari, Behnaz Shams, Azar Esmaeilzadeh, Narges Tanbakooei, Sara Koruji, Morteza Moghadam, Mojtaba Johari Ansari, Javad Mohajer Moghadam, Adel Johari Shabani, Ronak Stem Cell Res Ther Research BACKGROUND: Some children who have survived cancer will be azoospermic in the future. Performing isolation and purification procedures for spermatogonial stem cells (SSC) is very critical. In this regard, performing the process of decontamination of cancerous cells is the initial step. The major objective of the present study is to separate the malignant EL4 cell line in mice and spermatogonial stem cells in vitro. METHODS: The spermatogonial stem cells of sixty neonatal mice were isolated, and the procedure of co-culturing was carried out by EL4 which were classified into 2 major groups: (1) the control group (co-culture in a growth medium) and (2) the group of co-cultured cells which were separated using the microfluidic device. The percentage of cells was assessed using flow cytometry technique and common laboratory technique of immunocytochemistry and finally was confirmed through the laboratory technique of reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: The actual percentage of EL4 and SSC after isolation was collected at two outlets: the outputs for the smaller outlet were 0.12% for SSC and 42.14% for EL4, while in the larger outlet, the outputs were 80.38% for SSC and 0.32% for EL4; in the control group, the percentages of cells were 21.44% for SSC and 23.28% for EL4 (based on t test (p ≤ 0.05)). CONCLUSIONS: The present study demonstrates that the use of the microfluidic device is effective in separating cancer cells from spermatogonial stem cells. BioMed Central 2020-05-24 /pmc/articles/PMC7245899/ /pubmed/32448280 http://dx.doi.org/10.1186/s13287-020-01671-1 Text en © The Author(s) 2020 Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Ashtari, Behnaz
Shams, Azar
Esmaeilzadeh, Narges
Tanbakooei, Sara
Koruji, Morteza
Moghadam, Mojtaba Johari
Ansari, Javad Mohajer
Moghadam, Adel Johari
Shabani, Ronak
Separating mouse malignant cell line (EL4) from neonate spermatogonial stem cells utilizing microfluidic device in vitro
title Separating mouse malignant cell line (EL4) from neonate spermatogonial stem cells utilizing microfluidic device in vitro
title_full Separating mouse malignant cell line (EL4) from neonate spermatogonial stem cells utilizing microfluidic device in vitro
title_fullStr Separating mouse malignant cell line (EL4) from neonate spermatogonial stem cells utilizing microfluidic device in vitro
title_full_unstemmed Separating mouse malignant cell line (EL4) from neonate spermatogonial stem cells utilizing microfluidic device in vitro
title_short Separating mouse malignant cell line (EL4) from neonate spermatogonial stem cells utilizing microfluidic device in vitro
title_sort separating mouse malignant cell line (el4) from neonate spermatogonial stem cells utilizing microfluidic device in vitro
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7245899/
https://www.ncbi.nlm.nih.gov/pubmed/32448280
http://dx.doi.org/10.1186/s13287-020-01671-1
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