Transcriptome Sequencing and Comparative Analysis of Amphoteric ESCs and PGCs in Chicken (Gallus gallus).

Kai Jin,Yani Zhang,Jiuzhou Song,Qisheng Zuo,Guobing Chang,Bichun Li,Guohong Chen,Jing Zhou

doi:10.3390/ani10122228

Abstract

Simple SummaryThe study of chicken embryonic stem cells (ESCs) and primordial germ cells (PGCs) showed the potential application of developmental biology and translational medicine. However, the difference between amphoteric ESCs and PGCs is still elusive, limiting the accuracy of correlative research. In this paper, chicken amphoteric ESCs and PGCs were isolated, separated, and sequenced to explore their dynamic transcriptomes. Our results provide a knowledge base of transcriptomes in chicken amphoteric ESCs and PGCs, which will help other researchers interested in studying relative biological processes.Chicken (Gallus gallus) pluripotent embryonic stem cells (ESCs) and primordial germ cells (PGCs) can be broadly applied in the research of developmental and embryonic biology, but the difference between amphoteric ESCs and PGCs is still elusive. This study determined the sex of collected samples by identifying specific sex markers via polymerase chain reaction (PCR) and fluorescence activated cell sorter (FACS). RNA-seq was utilized to investigate the transcriptomic profile of amphoteric ESCs and PGCs in chicken. The results showed no significant differentially expressed genes (DEGs) in amphoteric ESCs and 227 DEGs exhibited in amphoteric PGCs. Moreover, those 227 DEGs were mainly enriched in 17 gene ontology (GO) terms and 27 pathways according to Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis. Furthermore, qRT-PCR was performed to verify RNA-seq results, and the results demonstrated that Notch1 was highly expressed in male PGCs. In summary, our results provided a knowledge base of chicken amphoteric ESCs and PGCs, which is helpful for future research in relevant biological processes.

Highlights

Chicken (Gallus gallus) is a classical model in developmental and embryonic biology [1].For decades, chickens have played a vital role in animal research as alternatives and outbredAnimals 2020, 10, 2228; doi:10.3390/ani10122228 www.mdpi.com/journal/animalsAnimals 2020, 10, 2228 experimental species to humans to compensate for ethical constraints [2,3,4]
For sample preparation for transcriptome analyses, the blastoderms and gonads were collected identified by polymerase chain reaction (PCR) chicken as described
No genes were differentially expressed in amphoteric embryonic stem cells (ESCs); differentially expressed genes were identified in amphoteric primordial germ cells (PGCs) and found to be highly enriched in significant

Summary

Introduction

Animals 2020, 10, 2228 experimental species to humans to compensate for ethical constraints [2,3,4] It works especially well when isolating and gaining a considerable number of pluripotent embryonic stem cells (ESCs) and primordial germ cells (PGCs) is much easier and convenient in chickens than other species due to the in ovo embryonic development [5,6]. This unique advantage accelerates the application of chicken. ESCs have been derived from cultures of chicken blastoderms taken from stage X-XII and PGCs can be obtained from the genital ridge (the precursor of gonad) at stage 28–30 embryos [6,11]

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