• 한국가축번식학회지
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• 제12권1호
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• pp.11-14
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• 1988

Utilizatin of primordial germ cell(PGC) as transferor of genetic materials is great potential in manipulating genes to promote genetic performances in chicken. This study explored that PGCs from early embryos as vehicle for molecular breeding strategles were isolated, these chromosomally marked donor PGCs were transplanted to germinal crescent of host embryos, and genetic materials of donor PGC were identified at the proliferative stage in host gonads.

• Asian-Australasian Journal of Animal Sciences
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• 제8권6호
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• pp.557-562
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• 1995

In this study, characteristics of chick primordial germ cells (PGCs), which is the founder cell of the germline, and gonadal development of the chick embryo between 12hrs and 6 day of incubation were investigated by transverse serial sections of chick embryos under the light microscopic observation. In embryo stage 20 (3 day of incubation), there are a lot of PGCs at the mesenchym, which were moving to the thickened epithelium (gonadal ridge). The PGCs arrive at both right and left gonad primordial in equal number prior to stage 24 (4 day of incubation), but in the following stages, the distribution of the PGCs became asymmetrical. More PGCs colonized the left than the right gonad, but the reason for the unequal distribution of PGCs is uncertain. The PGCs have mostly settled in the gonadal ridge (GR) at 6 day embryo. This study was conducted to investigate characteristics of the PGC migration and gonadal formation and observe the best condition for PGC isolation, culture and to attempt the possibility of the production for transgenic germline chimeras with manipulated PGCs.

• Molecules and Cells
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• 제38권10호
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• pp.895-903
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• 2015

Non-coding microRNAs (miRNAs) regulate the translation of target messenger RNAs (mRNAs) involved in the growth and development of a variety of cells, including primordial germ cells (PGCs) which play an essential role in germ cell development. However, the target mRNAs and the regulatory networks influenced by miRNAs in PGCs remain unclear. Here, we demonstrate a novel miRNAs control PGC development through targeting mRNAs involved in various cellular pathways. We reveal the PGC-enriched expression patterns of nine miRNAs, including miR-10b, -18a, -93, -106b, -126-3p, -127, -181a, -181b, and -301, using miRNA expression analysis along with mRNA microarray analysis in PGCs, embryonic gonads, and postnatal testes. These miRNAs are highly expressed in PGCs, as demonstrated by Northern blotting, miRNA in situ hybridization assay, and miRNA qPCR analysis. This integrative study utilizing mRNA microarray analysis and miRNA target prediction demonstrates the regulatory networks through which these miRNAs regulate their potential target genes during PGC development. The elucidated networks of miRNAs disclose a coordinated molecular mechanism by which these miRNAs regulate distinct cellular pathways in PGCs that determine germ cell development.

• Journal of Animal Science and Technology
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• 제55권5호
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• pp.417-425
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• 2013

동결 닭 PGCs의 생식계열 키메라를 이용한 생체에의 복원을 실용화 하기 위해서는, 닭 PGCs의 동결보존기술의 향상에 의해 동결 및 융해 후의 많은 생존세포를 확보 하는 것과, 생식계열 키메라의 제작효율을 높이는 것이 반드시 필요하다. 닭 PGCs는 배양 5.5일령의 닭 원시생식선으로부터 채취하고, ACS 방법에 의해서 순수 닭 PGCs를 분리했다. 닭 PGCs의 동결보존실험결과 다음의 4종류의 동결방법을 각각 비교 검토했다. 1. 플라스틱 스트로에 의한 완만동결법 (SF), 동결보호물질은 2M 에틸렌 글리콜 (EG), 2. 스트로에 의한 급속동결법 (RF), 8M EG + 7% PVP, 3. 동결용 Cryotube에 의한 SF, 2M EG, 4. 튜브에 의한 SF, 10% DMSO. 동결 및 융해 후의 PGCs의 생존율은 각각 76.4%, 70.6%, 80.5%, 78.1%로 관찰되었다.

• Asian-Australasian Journal of Animal Sciences
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• 제7권4호
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• pp.459-466
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• 1994

Primordial germ cells (PGCs) in aves are the progenitor cells for the gametes. These cells first appear in the epiblast (Eyal-Giladi et al.. 1981). Then translocate and concentrate to endoderm of germinal crescent area in the junction of the area opaca and area pellucida lateral to the primitive streak in stage 4 through 7. They separate from the endoderm, temporarily circulate via the blood vascular system, leave the blood vessels, and finally settle down in the gonadal anlagen at stage 20-24 where they rapidly proliferate to form germ cells. Recently, several attempts have been made to introduce foreign gene into the avian genome to form a transgenic chicken. The stem cells most readily available as vehicles for genetic manipulation of germline in avian species are the PGCs. PGCs have recently been manipulated genetically and used successfully as a vector for gene transfer.

• Asian-Australasian Journal of Animal Sciences
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• 제7권3호
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• pp.427-434
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• 1994

The primordial germ cells (PGCs) were transfected in vitro and expressed the exogenous RSVLTR/${\beta}G2$ plasmid, suggesting thaI PGC is a possible vector for direct gene transfer into the germ line. Transfection efficiency of cell suspensions containing PGCs was 1.5% by liposome mediated DNA transfection. By microinjection of the transfected PGCs into the host germinal crescent, PGCs migrated via blood vessel to the future gonad and these transfected PGCs resulted in the RSVLTR/${\beta}G2$ expression in the gonad. The results from the seeding of PGCs on the chorioallantoic membrane were insufficient to test the hypothesis that PGCs can penetrate or invade the chorioallantoic membrane for transport via the circulatory system.

• 한국발생생물학회:학술대회논문집
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• 한국발생생물학회 2003년도 제3회 국제심포지움 및 학술대회
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• pp.22-23
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• 2003

Primordial germ cell (PGC) is the progenitor cell of the germ cell lineage and eventually give rise to gametes that are responsible for creating individual organisms via a fertilization process. This means that PGC is a unique cell that can be converted into individual fish. This advantage of PGCs would make it possible to develop various applications in the field of fish bioengineering. First, PGCs may make it easier to preserve the genetic resources of fish. Cryopreservation of fish eggs or embryos has not been successfully achieved so far. Therefore, the only possible method to preserve genetic resources of fishes is to raise fish as live individuals. If PGCs isolated from various fishes could be cryopresewed, these cells could be converted into live fishes via germ-line chimera production. This is particularly useful for preserving genetic materials of endangered species. Even if the species of interest were to become extinct, it could be recovered by the transplantation of cryopreserved PGCs into the embryos of a closely related species. Another application of this technology is in what could be termed "surrogate broodstock technology". (중략)

• Asian-Australasian Journal of Animal Sciences
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• 제13권5호
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• pp.587-594
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• 2000

At present embryonic stem (ES) cells with confirmed pluripotential properties are only available in the mouse. Recently, we were able to isolate, culture and genetically transform primordial germ cell (PGC)-derived cells from pig embryos and demonstrate their ability to contribute to chimera development in the pig. In order to determine whether the system we developed could be used to isolate embryonic germ (EG) cells from other mammalian species, we placed isolated PGCs from cattle, goats, rabbits and rats in culture. Briefly, PGCs were isolated from fetuses of cow (day 30-50), goat (day 25), rabbit (day 15-18) and rat (day 11-12), and plated on STO feeder cells in Dulbecco's modified Eagle's medium (DMEM): Ham's F10 medium (1:1) supplemented with 0.01 mM nonessential amino acids, 2 mM L-glutamine, 0.1 mM $\beta$ - mercaptoethnol, soluble recombinant human stem cell factor (SCF; 40ng/ml), human basic fibroblast growth factor (bFGF; 20ng/ml) and human leukemia inhibitory factor (LIF; 20ng/ml). For maintenance of the cells, colonies were passed to fresh feeders every 7-10 days. In all species tested, we were able to obtain and maintain colonies with ES-like morphology. Their developmental potential was tested by alkaline phosphatase (AP) staining and in vitro differentiation assay. For genetic transformation, cells were electroporated with a construct containing the green fluorescent protein (GFP) under the control of the cytomegalovirus (CMV) promoter. GFP-expressing colonies were detected in cattle, rabbits and rats. These results suggest that PGC-derived cells from cattle, goats, rabbits and rats can be isolated, cultured, and genetically transformed, and provide the basis for analyzing their developmental potential and their possible use for the precise genetic modification of these species.

• 한국동물학회지
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• 제27권1호
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• pp.13-24
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• 1984

산개구리의 germ plasm 및 축결정 세포질에 미치는 자외선의 영향을 수량적, 시간적으로 조사하였다. 산개구리 난의 식물극을 수정 후 60분경에 1,600 $ergs/mm^2$이상의 자외선을 조사하였을때 40%이상의 시원생식세포의 수가 감소하였으나, 그 이하의 양에서는 전혀 영향이 없었다. 그러나 1,600 $ergs/mm^2$부터는 시원생식세포 수의 감소 정도는 자외선의 양에 비례하였다. 또 신경관 형성에 전혀 영향을 미치지 않으면서 시원생식세포의 형성을 완전히 억제시키는 자외선의 양은 4,800 $ergs/mm^2$이었다. 이에 4,800 $ergs/mm^2$의 자외선을 수정 후 일정한 시간간격으로 조사하여 germ plasm에 미치는 자외선의 영향을 조사하였다. 그 결과 자외선이 조사된 시기가 이를수록 시원생식세포수의 감소가 크며, 수정후 60분부터 차츰 감소율이 줄어 들다가 3분열 이후에는 자외선의 효과가 거의 없음을 관찰하였다. 또한 시원생식세포와 축형성에 미치는 자외선의 영향을 시기적으로 비교한 결과 산개구리난의 축결정이 다른 무미양서류에서와 같이 수정에서 제1분열 사이의 $0.7\\sim0.8$시기 이전에 이루어지므로 그 이후에는 자외선의 감응도가 급격히 저하되었다. 그러나 germ plasm의 자외선에 대한 감응도는 훨씬 늦게까지 지속되어 8구기에 이르기까지 그 영향이 나타났다.

• 한국가금학회:학술대회논문집
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• 한국가금학회 2001년도 제18차 정기총회 및 학술발표 PROCEEDINGS
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• pp.69-70
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• 2001

Lectins have great potential as to determine the alternation of the distribution of cell surface carbohydrates during cellular development and differentiation. Here, we investigated the presence and distribution of cell surface carbohydrates on chicken primordial germ cells (PGCs) during the migration and gonadal stages using a variety of lectins. A total of six FITC-labelled lectins from several specificity classes were used: ConA (glucose/mannose), WGA (N-acetylglucosamine), STA (N-acetylglucosamine), DBA (N-acetylgalactosamine/galactose), UEA-I (fucose) and PHA-E (oilgosaccharide). As a results, PGC-specific binding was observed in STA. PGCs of migration stage (2.5- and 5.5-day embyos) were STA-positive whereas PGCs of 10-day embryonic gonad were not. The results suggest that N-acetylglucosamine residuse are present specifically in migrating chicken PGCs and changes during development.