TY - JOUR

T1 - Establishment of the Primary Avian Gonadal Somatic Cell Lines for Cytogenetic Studies

AU - Pristyazhnyuk, Inna E.

AU - Malinovskaya, Lyubov P.

AU - Borodin, Pavel M.

N1 - Funding Information: Funding: Data collection and analysis was funded by Russian Science Foundation, grant number 20-64-46021. Access to microscopic equipment and xCELLigence instrument was granted by the Core Facility for Microscopy of Biologic Objects of Siberian Branch of Russian Academy of Sciences, Novosibirsk, Russia (regulation no. 3054) funded by the Ministry of Science and Higher Education of the Russian Federation, grant number FWNR-2022-0019. Open access was supported by Ministry of Science and Higher Education of the Russian Federation, grant number #2019-0546 (FSUS-2020-0040). Publisher Copyright: © 2022 by the authors. Licensee MDPI, Basel, Switzerland.

PY - 2022/7/1

Y1 - 2022/7/1

N2 - 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.

AB - 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.

KW - avian primary cell culture

KW - electroporation

KW - karyotyping

KW - songbirds

UR - http://www.scopus.com/inward/record.url?scp=85133252664&partnerID=8YFLogxK

U2 - 10.3390/ani12131724

DO - 10.3390/ani12131724

M3 - Article

C2 - 35804624

AN - SCOPUS:85133252664

VL - 12

JO - Animals

JF - Animals

SN - 2076-2615

IS - 13

M1 - 1724

ER -