Zebra finch PGCs transcriptome
Primordial germ cells (PGCs) are the stem cells precursors of spermatocytes and oocytes. In avian species they can be found in the blood of early embryos during their migration process to the gonadal anlage. Interestingly, cultured and genetically modified PGCs can be used to generate transgenic birds after being reinjected into surrogate embryos. However, many factors and variables can play a role to maintain and expand those cells in vitro. Herewith, In order to elucidate the effect of culture conditions and time on the gene expression, PGCs were freshly extracted from embryonic blood gained from zebra finch male embryos (14HH), cultured for one day (G1) and for 10 days (G10DIV) in a specific culture media. The RNAseq data for the two conditions, were generated from RNAseq libraries of 4 replicates, each containing 400 cells. The first strand cDNA synthesis and PCR amplification were carried out according to the protocol of SMART Seq v4 Ultra Low Input RNA Kit (TAKARA). Tagmentation and indexing of the amplified cDNAs were performed using the Nextera XT library preparation kit (Illumina) with multiplexing primers, according to protocol provided by the manufacturer. Differences in the gene expression profile and pathways related to proliferation and survival could give new insights regarding the growth factors required to extend the survival time and increase the proliferation rate of PGCs in vitro.