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Preventing Microbial Contamination during Long-Term In Vitro Culture of Human Granulosa-Lutein Cells: An Ultrastructural Analysis

Purpose. To investigate whether the addition of antibiotic/antimycotic during human granulosa-lutein cells (GLCs) isolation and cell-plating procedures prevents microbial contamination after 144 h of culture and also evaluate the effects of contamination on GLCs ultrastructure and steroid secretion....

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Autores principales: Campos, C. O., Bernuci, M. P., Vireque, A. A., Campos, J. R., Silva-de-Sá, M. F., Jamur, M. C., Rosa-e-Silva, A. C. J. S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: International Scholarly Research Network 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3439989/
https://www.ncbi.nlm.nih.gov/pubmed/22988519
http://dx.doi.org/10.5402/2012/152781
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author Campos, C. O.
Bernuci, M. P.
Vireque, A. A.
Campos, J. R.
Silva-de-Sá, M. F.
Jamur, M. C.
Rosa-e-Silva, A. C. J. S.
author_facet Campos, C. O.
Bernuci, M. P.
Vireque, A. A.
Campos, J. R.
Silva-de-Sá, M. F.
Jamur, M. C.
Rosa-e-Silva, A. C. J. S.
author_sort Campos, C. O.
collection PubMed
description Purpose. To investigate whether the addition of antibiotic/antimycotic during human granulosa-lutein cells (GLCs) isolation and cell-plating procedures prevents microbial contamination after 144 h of culture and also evaluate the effects of contamination on GLCs ultrastructure and steroid secretion. Methods. GLCs obtained from five women submitted to assisted reproductive techniques (ARTs) were isolated with PBS supplemented with antibiotic/antimycotic or PBS nonsupplemented and cultured for 144 h. GLCs were evaluated by transmission electron microscopy (TEM), and estradiol (E2) and progesterone (P4) secretion was assayed by chemiluminescence. Results. Although no contaminating microorganisms were identified by light microscopy, TEM analyses revealed several bacterial colonies in culture dishes of GLCs isolated with only PBS. Bacterial contamination disrupted the adherence of the GLCs to the culture plate interfering with monolayer formation affecting the growth pattern of GLCs. Various cellular debris and bacteria were observed, and no organelles were found in the cytoplasm of infected cells. While bacterial contamination decreased estradiol media levels, it increased progesterone, as compared with noncontaminated group. Conclusion. Taken together, our data showed that the addition of a high dose of antibiotic/antimycotic during the isolation and cell-plating procedures prevents microbial contamination of long-term GLCs culture as its effects on cells growth and function in vitro.
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spelling pubmed-34399892012-09-17 Preventing Microbial Contamination during Long-Term In Vitro Culture of Human Granulosa-Lutein Cells: An Ultrastructural Analysis Campos, C. O. Bernuci, M. P. Vireque, A. A. Campos, J. R. Silva-de-Sá, M. F. Jamur, M. C. Rosa-e-Silva, A. C. J. S. ISRN Obstet Gynecol Clinical Study Purpose. To investigate whether the addition of antibiotic/antimycotic during human granulosa-lutein cells (GLCs) isolation and cell-plating procedures prevents microbial contamination after 144 h of culture and also evaluate the effects of contamination on GLCs ultrastructure and steroid secretion. Methods. GLCs obtained from five women submitted to assisted reproductive techniques (ARTs) were isolated with PBS supplemented with antibiotic/antimycotic or PBS nonsupplemented and cultured for 144 h. GLCs were evaluated by transmission electron microscopy (TEM), and estradiol (E2) and progesterone (P4) secretion was assayed by chemiluminescence. Results. Although no contaminating microorganisms were identified by light microscopy, TEM analyses revealed several bacterial colonies in culture dishes of GLCs isolated with only PBS. Bacterial contamination disrupted the adherence of the GLCs to the culture plate interfering with monolayer formation affecting the growth pattern of GLCs. Various cellular debris and bacteria were observed, and no organelles were found in the cytoplasm of infected cells. While bacterial contamination decreased estradiol media levels, it increased progesterone, as compared with noncontaminated group. Conclusion. Taken together, our data showed that the addition of a high dose of antibiotic/antimycotic during the isolation and cell-plating procedures prevents microbial contamination of long-term GLCs culture as its effects on cells growth and function in vitro. International Scholarly Research Network 2012-09-03 /pmc/articles/PMC3439989/ /pubmed/22988519 http://dx.doi.org/10.5402/2012/152781 Text en Copyright © 2012 C. O. Campos et al. https://creativecommons.org/licenses/by/3.0/ This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Clinical Study
Campos, C. O.
Bernuci, M. P.
Vireque, A. A.
Campos, J. R.
Silva-de-Sá, M. F.
Jamur, M. C.
Rosa-e-Silva, A. C. J. S.
Preventing Microbial Contamination during Long-Term In Vitro Culture of Human Granulosa-Lutein Cells: An Ultrastructural Analysis
title Preventing Microbial Contamination during Long-Term In Vitro Culture of Human Granulosa-Lutein Cells: An Ultrastructural Analysis
title_full Preventing Microbial Contamination during Long-Term In Vitro Culture of Human Granulosa-Lutein Cells: An Ultrastructural Analysis
title_fullStr Preventing Microbial Contamination during Long-Term In Vitro Culture of Human Granulosa-Lutein Cells: An Ultrastructural Analysis
title_full_unstemmed Preventing Microbial Contamination during Long-Term In Vitro Culture of Human Granulosa-Lutein Cells: An Ultrastructural Analysis
title_short Preventing Microbial Contamination during Long-Term In Vitro Culture of Human Granulosa-Lutein Cells: An Ultrastructural Analysis
title_sort preventing microbial contamination during long-term in vitro culture of human granulosa-lutein cells: an ultrastructural analysis
topic Clinical Study
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3439989/
https://www.ncbi.nlm.nih.gov/pubmed/22988519
http://dx.doi.org/10.5402/2012/152781
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