Project description:The present investigation was to identify the signaling and metabolic pathways of expressed genes by microarray comparison between Primordial Germ Cells (PGCs) and their somatic counterpart, chicken embryonic fibroblasts (CEFs). We identified a total of 2,605 expressed transcripts. Among these, 1,197 were predominantly expressed in PGCs, and 1,408 were predominantly expressed in CEFs.
Project description:The present investigation was to identify the signaling and metabolic pathways of expressed genes by microarray comparison between Primordial Germ Cells (PGCs) and their somatic counterpart, chicken embryonic fibroblasts (CEFs). We identified a total of 2,605 expressed transcripts. Among these, 1,197 were predominantly expressed in PGCs, and 1,408 were predominantly expressed in CEFs. Cell culture and microarray data generation were performed in triplicate. Blood PGCs from E2.5 embryos (N=20 for each replication) and CEFs from E6.5 embryos (N=5 for each replication) were cultured in appropriate culture medium. Total RNA was extracted from cultured PGCs (1,500,000 cells for each replication) and cultured CEFs (6,000,000 cells for each replication) with a Qiagen RNeasy kit. About 5 M-BM-5g of total RNA from each replication was used for labeling. Probe synthesis from total RNA samples, hybridization, detection, and scanning were performed according to standard protocols from Affymetrix.
Project description:The aim of this investigation was to identify differentially expressed transcripts in chicken primordial germ cells (PGCs) compared with blastoderms, chicken embryonic fibroblasts (CEFs) and gonadal stromal cells (GSCs). Based on the P values (P < 0.05) and fold-change cutoff (4.0), several transcripts were differentially expressed in PGCs. However, we restricted our analysis to identify gene-matched transcripts only. Finally, we identified 1,505 differentially expressed transcripts in PGCs compared with blastoderms, 760 differentially expressed transcripts in PGCs compared with CEFs, and 466 differentially expressed transcripts in PGCs compared with GSCs.
Project description:The comparative transcriptomic analysis of chicken stem cells defines a chicken set of pluripotency associated genes that is almost coincident with mammalian pluripotency-assocaited genes. A total of 5944 differentially expressed unique identifiers (target IDs) were defined for CEF, 5942 for BM2 cells, 4550 for cES, 5465 for PGC and 5522 for cBC. Chicken Blastodermal cells (cBC) were taken from stage IX-XII (Eyal-Giladi & Kochav, 1976) embryos,chicken embryonic stem (cES) cells were established, amplified on inactivated STO feeder cells in proliferative medium containing cytokines and growth factors as described (Pain et al., 1996, Lavial et al., 2007). Long term cultured primordial germ cells (PGCs) were derived from 48h embryonic blood and maintained as described (McDonald et al., 2010). The non-tumorigenic BM2 monocytic cell line was grown as described (Solari et al., 1996). Primary chicken Embryonic fibroblasts (CEF) were prepared from 11-12 day old embryos according to standard protocols (Gandrillon et al., 1987), maintained and homogenised for 3-4 passages before being used as a somatic cell control. RNAs were extracted using Trizol (Invitrogen) according to the manufacturer and microarray analysis performed in biological triplicate.
