Survival and Proliferation of Refined Chicken Circulating Primordial Germ Cells Cultured In Vitro

Abstract

Abstract. In recent years, much attention has been paid to the manipulation of primordial germ cells (PGCs) to produce transgenic chicken. For this purpose, the in vitro proliferation of PGCs would make gene targeting possible, promoting the efficiency of transgenes. However, few researches have been carried out on this topic. In the present study, we tried to culture chicken PGCs outside the body. The PGCs were isolated from circulating blood of chicken embryos by the method of Ficoll density centrifugation, and labeled with PKH26 red fluorescent cell linker. The cells were refined by selection with a fine glass pipette and a micromanipulator in order to avoid the contamination of PGCs with red cells. The refined cells were then cultured for a given period. The PGCs proliferated when they were cultured on feeder cells derived from the germinal ridges of chicken embryos at stage 27. During a 5 day culture period, the highest rate of increased PGCs was about 51% for one of the two test groups. Although some kinds of growth factors were supplemented in the culture media, no synergistic effect of chicken stem cell factor (chSCF) with murine leukemia inhibitory factor (LIF) and human basic fibroblast growth factor (bFGF) on the proliferation of PGCs was found. This result implies that the LIF and bFGF influencing the proliferation of PGCs may not be conserved between mammals and birds.

Chicken stem cell factor enhances primordial germ cell proliferation cooperatively with fibroblast growth factor 2

“…Our data showed that chSCF2-BRL feeder cells improved the in vitro proliferation rate of chicken PGCs by fivefold compared with normal BRL feeder cells. Meanwhile, our data also demonstrated that chSCF1-BRL feeder cells did not sufficiently supported the in vitro proliferation of chicken PGCs, and this result was consistent with the previous studies [ 17 , 33 ]. Thus, chSCF2 was considered one of the essential factor for the in vitro proliferation of chicken PGCs.…”

Long-Term Proliferation of Chicken Primordial Germ Cells Cultured In Vitro.

“…Our previous study suggested that some of growth factors such as LIF and bFGF might have influenced the proliferation of chicken circulating PGCs in vitro [13,14]. In the present experiment, therefore, we tried A of the area pellucida of developing embryos [1,2].…”