Establishment of the Primary Avian Gonadal Somatic Cell Lines for Cytogenetic Studies.

Autor: Pristyazhnyuk IE; Department of Molecular Genetics, Cell Biology and Bioinformatics, Institute of Cytology and Genetics, Siberian Department of Russian Academy of Sciences, 630090 Novosibirsk, Russia., Malinovskaya LP; Department of Cytology and Genetics, Faculty of Natural Sciences, Novosibirsk State University, 630090 Novosibirsk, Russia., Borodin PM; Department of Molecular Genetics, Cell Biology and Bioinformatics, Institute of Cytology and Genetics, Siberian Department of Russian Academy of Sciences, 630090 Novosibirsk, Russia.
Jazyk: angličtina
Zdroj: Animals : an open access journal from MDPI [Animals (Basel)] 2022 Jul 04; Vol. 12 (13). Date of Electronic Publication: 2022 Jul 04.
DOI: 10.3390/ani12131724
Abstrakt: 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.
Databáze: MEDLINE
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