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

Abstract

Simple SummaryWe developed a simple method for primary somatic cell culture establishment from the ovaries of the great tits and testes of ten Passerine species. The ovary-derived cell cultures were cultivated until the tenth passage without any noticeable decrease in their proliferative activity, while testis-derived cell cultures demonstrated a decreased proliferation potential. However, sufficient material was available from both cell cultures originating from the ovary and testis to make excellent mitotic metaphase chromosomal preparations. We demonstrated the high efficiency of electroporation for genetic modification of the ovary-derived cell line. Thus, the established ovary-derived cell line could be efficiently used in cytogenetic and genomic 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) 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.