Establishment of the Primary Avian Gonadal Somatic Cell Lines for Cytogenetic Studies
The last decade was marked by a steep rise in avian studies at genomic and cellular levels. Cell lines are important tools for in vitro studies in cell biology and cytogenetics. We developed a simple method of primary somatic cell culture establishment from the ovaries of the great tits (Parus major...
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ftmdpi:oai:mdpi.com:/2076-2615/12/13/1724/ 2023-08-20T04:05:23+02:00 Establishment of the Primary Avian Gonadal Somatic Cell Lines for Cytogenetic Studies Inna E. Pristyazhnyuk Lyubov P. Malinovskaya Pavel M. Borodin agris 2022-07-04 application/pdf https://doi.org/10.3390/ani12131724 EN eng Multidisciplinary Digital Publishing Institute Animal Genetics and Genomics https://dx.doi.org/10.3390/ani12131724 https://creativecommons.org/licenses/by/4.0/ Animals; Volume 12; Issue 13; Pages: 1724 avian primary cell culture songbirds karyotyping electroporation Text 2022 ftmdpi https://doi.org/10.3390/ani12131724 2023-08-01T05:36:20Z The last decade was marked by a steep rise in avian studies at genomic and cellular levels. Cell lines are important tools for in vitro studies in cell biology and cytogenetics. We developed a simple method of primary somatic cell culture establishment from the ovaries of the great tits (Parus major) and testes of ten Passerine species, characterized the cellular composition of the ovary-derived lines using RT-PCR and immunolocalization of the tissue-specific markers and tested the efficiency of two methods of genetic transformation of the ovary-derived cell line. We found that the ovary-derived cell cultures of the great tit were composed of fibroblasts mainly, but also contained interstitial and granulosa cells. They were cultivated until the 10th passage without any noticeable decrease in their proliferative activity. The testis-derived cell cultures had lower proliferative potential. However, both ovary- and testis-derived cell cultures provided enough material for high quality mitotic metaphase chromosome preparations. The efficiency of its transduction with lentivirus containing a GFP reporter was very low, while electroporation with episomal vectors expressing GFP resulted in a high yield of GFP-positive cells. The proposed method could be used for the generation of high quality material for various cytogenetic and genomic studies. Text Avian Studies MDPI Open Access Publishing Parus ENVELOPE(3.950,3.950,-71.983,-71.983) Animals 12 13 1724 |
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MDPI Open Access Publishing |
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language |
English |
topic |
avian primary cell culture songbirds karyotyping electroporation |
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avian primary cell culture songbirds karyotyping electroporation Inna E. Pristyazhnyuk Lyubov P. Malinovskaya Pavel M. Borodin Establishment of the Primary Avian Gonadal Somatic Cell Lines for Cytogenetic Studies |
topic_facet |
avian primary cell culture songbirds karyotyping electroporation |
description |
The last decade was marked by a steep rise in avian studies at genomic and cellular levels. Cell lines are important tools for in vitro studies in cell biology and cytogenetics. We developed a simple method of primary somatic cell culture establishment from the ovaries of the great tits (Parus major) and testes of ten Passerine species, characterized the cellular composition of the ovary-derived lines using RT-PCR and immunolocalization of the tissue-specific markers and tested the efficiency of two methods of genetic transformation of the ovary-derived cell line. We found that the ovary-derived cell cultures of the great tit were composed of fibroblasts mainly, but also contained interstitial and granulosa cells. They were cultivated until the 10th passage without any noticeable decrease in their proliferative activity. The testis-derived cell cultures had lower proliferative potential. However, both ovary- and testis-derived cell cultures provided enough material for high quality mitotic metaphase chromosome preparations. The efficiency of its transduction with lentivirus containing a GFP reporter was very low, while electroporation with episomal vectors expressing GFP resulted in a high yield of GFP-positive cells. The proposed method could be used for the generation of high quality material for various cytogenetic and genomic studies. |
format |
Text |
author |
Inna E. Pristyazhnyuk Lyubov P. Malinovskaya Pavel M. Borodin |
author_facet |
Inna E. Pristyazhnyuk Lyubov P. Malinovskaya Pavel M. Borodin |
author_sort |
Inna E. Pristyazhnyuk |
title |
Establishment of the Primary Avian Gonadal Somatic Cell Lines for Cytogenetic Studies |
title_short |
Establishment of the Primary Avian Gonadal Somatic Cell Lines for Cytogenetic Studies |
title_full |
Establishment of the Primary Avian Gonadal Somatic Cell Lines for Cytogenetic Studies |
title_fullStr |
Establishment of the Primary Avian Gonadal Somatic Cell Lines for Cytogenetic Studies |
title_full_unstemmed |
Establishment of the Primary Avian Gonadal Somatic Cell Lines for Cytogenetic Studies |
title_sort |
establishment of the primary avian gonadal somatic cell lines for cytogenetic studies |
publisher |
Multidisciplinary Digital Publishing Institute |
publishDate |
2022 |
url |
https://doi.org/10.3390/ani12131724 |
op_coverage |
agris |
long_lat |
ENVELOPE(3.950,3.950,-71.983,-71.983) |
geographic |
Parus |
geographic_facet |
Parus |
genre |
Avian Studies |
genre_facet |
Avian Studies |
op_source |
Animals; Volume 12; Issue 13; Pages: 1724 |
op_relation |
Animal Genetics and Genomics https://dx.doi.org/10.3390/ani12131724 |
op_rights |
https://creativecommons.org/licenses/by/4.0/ |
op_doi |
https://doi.org/10.3390/ani12131724 |
container_title |
Animals |
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12 |
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13 |
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1724 |
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1774715894632546304 |