Isolation and Number of Gonadal Primordial Germ Cells (Gonadal PGCs) on the Stages of Early Embryonic Development of KUB Chicken

Abstract

Primordial germ cells (PGCs) are cells that will differentiate themselves into spermatogonia in the testis or oogonia in the ovary. Primordial germ cells arise from epiblast and circulate through the bloodstream and finally entering gonadal anlage. The aim of this study was to determine the number of gonadal PGCs of KUB chicken at different development stages. Sixty KUB chicken fertile eggs were divided into four groups (6, 7, 8, and 9 days incubation periods), and incubated at 38 o C with a humidity of 60%. Harvesting was synchronized to the embryonic development at 6-9 d. Gonads were collected using sharp tweezers, and were placed in Eppendorf tube 1.5 mL containing 500 µL PBS [-]. Gonadal PGCs were purified using PBS [-]. The results showed that the average number of gonadal PGCs at 6, 7, 8, and 9 d were 113.7; 143.5; 92.9; and 85.7 cells per embryo, respectively. Number of gonadal PGCs per embryo of KUB chicken were significantly affected by stage of embryonic development (P<0.05), which reached a peak at day 7 of incubation, so that the isolation and collection of PGCs from the gonads were recommended at day 7 of incubation. This information is useful in production of germline chimera of other Indonesian local chickens. Keywords

Dynamics of the transcriptome during chicken embryo development based on primordial germ cells

“…In contrast, there is limited information on the regulation of gene expression during the migration of PGCs in vivo. This is mainly because of the limited number of available PGCs [ 20 ]. The new technique for isolating viable avian PGCs combined with the method of RNA isolation from a single cell offers a unique approach for the study of cell transcriptome dynamics.…”

Comparison of the Transcriptomic and Epigenetic Profiles of Gonadal Primordial Germ Cells of White Leghorn and Green-Legged Partridgelike Chicken Embryos

“…The results of Szczerba et al [ 19 ] showed that the number of gPGCs was probably influenced by the stage of embryonic development. In the case of inbred lines or endangered breeds, the number of total PGCs seems to be lower compared to commercial chicken breeds [ 23 , 24 ]. This lower number of PGCs seems likely to be correlated with the lower productivity of the local chicken.…”

Panjang Gonad Dan Jumlah Primordial Germ Cell Ayam White Leghorn

“…Perbedaan ini kemungkinan disebabkan oleh perbedaan metode pemurnian sel dan perkembangan umur embiro. Jumlah PGC-gonad dipengaruhi oleh perkembangan embiro, dimana jumlah PGC-gonad secara signifikan meningkat dan mencapai puncaknya pada hari ke-7, karena PGC sebagian besar telah menetap di bagian gonad dan setelah itu jumlah PGC akan menurun (Sopiyana et al, 2017).…”

“…Namun, dibandingkan dengan perolehan jumlah PGC-gonad ayam KUB pada umur embrio yang sama hanya mendapatkan jumlah PGC-gonad sebesar 113,7 sel per embrio (Sopiyana et al, 2017). Sementara itu, jumlah PGC-gonad ayam KUB yang dimurnikan dengan metode PBS (-) dengan waktu inkubasi embrio dalam PBS (-) selama 1 jam yaitu 151,47 sel per embrio pada umur embrio 7 hari (Kostaman dan Sopiyana, 2017).…”

“…Primordial germ cell (PGC)-gonad pada ayam mempunyai karakteristik morfologi, yaitu memiliki ukuran sel yang besar (12,73-25,11 µm), bentuk bulat dengan kontur tidak teratur, inti yang sperikal dan besar yang letaknya tidak simetris. Selain itu, PGC-gonad mempunyai berat jenis sebesar 1.029-1.040 (Sopiyana et al, 2017).…”