• Proceedings of the Korea Society of Poultry Science Conference
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• 2001.11a
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• pp.71-73
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• 2001

In this study, we could improve transmission efficiency of germline chimeras by transfer of gonadal PGCs (gPGCs) cultured in vitro. Of hatched recipient chicks, 301 chickens (141 males and 160 females) were brought up to sexual maturity and these WLs (KOC) were mated with KOCs for testcross, resulting in 27 germline chimeras (15 males and 12 females) identified by black feather color of their progenies. The production efficiency of germline chimera production of experimental groups was observed (P=0.6831). The average transmission efficiency of proven germline chimeras was 0.6 ∼56.5% (15.0% on average). The transmission efficiency of experimental group which were transferred 10-days cultured gPGCs without Ficoll treatment was highest (49.7%) and that of experimental stock which transferred non-cultured gPGCs with Ficoll treatment was lowest (0.6%). The duration of in vitro culture before transferring was significantly important for the high efficiency of germline transmission. Transferring 10-days cultured gPGCs made the transmission efficiency higher rather than transferring non-cultured and 5-days cultured gPGCs, 50 times and 10 times, respectively (p<0.0001). However, Ficoll treatment for increasing the population ratio of gPGCs negatively affected the transmission efficiency and the effects of sexuality and the reciprocal interaction between treatments showed no significant differences. These findings demonstrated that the crucial factors for improving the germline transmission were the duration of in vitro culture prior to transfer. Thus, we developed the complete system for production of germline chimera using cultured gPGCs with highly improved efficiency and this system would be useful for genetic manipulation and obtaining the transgenic aves.

• Korean Journal of Poultry Science
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• v.41 no.4
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• pp.249-259
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• 2014

Cryopreserving cells which are maintaining their viability are the very complex process. This study has been carried out in order to find the effects of cryopreservation steps and freezing media on the rates of viability of cryopreserved chicken primordial germ cells (PGCs). PGCs obtained from the germinal gonade of 5.5~6 day (stage 28) chick embryos of Korean Ogye (KO) and Commercial breeds (C), using the MACS method were suspended in a freezing medium containing a freezing and protecting agents (e.g. dimethyl sulfoxide (DMSO), ethylene glycol (EG) and propylene glycol (PG)). Gonads were harvested from stage 28 chick embryos and pooled in groups of 5, 10, 15, 20E embryos, contributing gonads to the cell suspension. The gonadal cells, including PGCs, were then frozen in 1 of the following cryoprotectant treatments : 2.5%, 5%, 10%, 15% and 0% cryoprotectant (DMSO, EG, PG) as a control. Effects of exposure to slow freezing and vitrification, with different concentrations of the cryoprotectant solution, were examined. After vitrification and slow freezing, survival rates of the frozen-thawed PGCs from the 10% EG plus FBS treatment were 85.63%, and 66.14% (p<0.05), respectively. The viability of PGCs after freeze-thawing was significantly higher for 10% EG plus FBS treatment than for 10% PG + FBS treatment (p<0.05) (85.63% vs 66.81%) by vitrification. This study established a method for preserving chicken PGCs that enables systematic storage and labeling of cryopreserved PGCs in liquid ($LN_2$) at a germplasm repository and ease of entry into a data base. In the future, the importance for this new technology is that poultry lines can be conserved while work is being conducted on improving the production of germline chimeras.

• KOREAN POULTRY JOURNAL
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• v.52 no.4
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• pp.144-146
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• 2020

• Korean Journal of Ichthyology
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• v.13 no.4
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• pp.248-253
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• 2001

An histological study was conducted to determine the initial treatment time and treatment duration in the use of sex-reversal hormones in relation to gonadal development and sexual differentiation in the bagrid catfish, Leiocassis ussuriensis. The primordial germ cell, which could be recognized from one-day-old fry, began to protrude into the peritoneal cavity between the mesonephric duct and the gut. The primordial gonad with a genital ridge was developed at 5~10 days after hatching. Sex differentiation of the ovary was identified by the ovarian cavity and meiotic oocytes from 20-day-old larvae. Testicular differentiation was also identified by spermatogonial cells from 20-day-old larvae. It may therefore be concluded that this species belongs to the differentiated type of gonochoristic teleost.

• Development and Reproduction
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• v.4 no.2
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• pp.137-145
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• 2000

The regulatory mechanisms of the initiation and the formation of ovarian follicles during fetal stage of mammals are largely unknown. In addition to the gonadotropins secreted from pituitary, various growth factors, and steroid hormones are believed to be involved in the differentiation and initiation of growth of primordial follicles consisting of primordial germ cells migrated from yolk sac and streamed cells from mesonephric somatic cells. In human, primordial follicles that have already initiated differentiation at fetal stage undergo either folliculogenesis to ovulate or atresia after growth. Some of primordial follicles remain without growth for 50 years or longer. The objective of this paper is to review the mechanism of the formation, growth arrest, and initiation of primordial follicles in human fetal and neonatal ovaries.

• Proceedings of the Korea Society of Poultry Science Conference
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• 2001.11a
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• pp.74-76
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• 2001

Comparing to mammals, male bird has the homozygote ZZ and female has the heterozygote n. Therefore, the sex of fertilized eggs is defined by female chromosome constitution. Although this cytological observation had been established, the molecular and cellular mechanism of germ cell differentiation are essentially unknown in aves. Especially, the differentiation of germ cells in mixed-sex chimeras has not yet been clearly elucidated. Primordial germ cells, which are the progenitors of sperm or egg after sexual maturity, firstly arise in the epiblast and migrate to embryonic gonads through the blood vessel. During the embryo development, these PGCs differentiate in the pathway of mate or female, respectively and develop the sperm or egg cells after sexual maturity. In this paper, we confirmed that the female PGCs could migrate into the recipient male gonads after transferring and differentiate into germ cells in the embryonic stages. The primordial germ cells were isolated from the female embryonic gonads of 5.5-day-old incubation and re-injected into the male recipient embryos of 2-day-old incubation, which produced mixed-sex chimera in the germline. The finding in this study demonstrated the ability of migration and differentiation of gonadal primordial germ cells in mixed-sex chicken.

• Development and Reproduction
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• v.10 no.3
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• pp.193-196
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• 2006

The process in the formation of primordial germ cells, primitive and early gonadogenesis were investigated by histological examination for the reproductive physiological study in the scorpion fish, Sebastiscus marmoratus. The primordial germ cells about $10\;{\mu}m$ in diameter were observed in the fibrous mesenchymal tissue located between gut and mesonephric duct of the larvae within the maternal body. As the fibrous epithelium extends to the direction of peritoneum, the primordial germ cells moved. From 31 days post parturition, the fibrous epithelium gets thicker, and the primitive gonad starts to form. From 49 days post parturition, as the primitive gonad of the primitive gonad extend, it starts to form cavity, and about 60 days post parturition, the formation of the ovarian cavity is completed. From 79 days post parturition, germ cells proliferated along the inner edge of the ovarian cavity.

• Journal of Aquaculture
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• v.18 no.2
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• pp.76-80
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• 2005

The early gonadal development and sex differentiation of the mandarin fish, Siniperca scherzeri was described from hatch to the 130th day post-hatch (DPH). Primordial germ cells were observed on the just hatched fry (5.10 mm in total length (TL)) and began to protrude into the peritoneal cavity between mesonephric duct and gut. The primordial gonad, with the formation of genital ridge, developed on the 8-10th DPH (10.77-12.47 mm TL). Ovarian differentiation was identified by the presence of ovarian cavity and meiotic oocytes from the 40th DPH (29.75 mm TL). Testicular differentiation was identified by the presence of spermatogonial cells with efferent duct also from the 40th DPH. Sex differentiation of the mandarin fish was identified as gonochoristic type.

• Korean Journal of Poultry Science
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• v.40 no.3
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• pp.197-205
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• 2013

This study established a method for preserving chicken primordial germ cells (PGCs) that enables long-term storage in liquid nitrogen ($LN_2$) for preservation of the species. The purpose of this study was to compare the effects of Ethylene Glycol (EG) and Propylene Glycol (PG) on viability of cryopreserved PGCs with vitrification in Korean Native Chicken (Ogye), and to fine should be find or to the optimal protocol for PGCs freezing. One of the important components of cryopreservation process is cryopreservation medium that plays a vital role in preventing cellular injury during freeze-thawing. Cryoprotective agents have been known to improve cell viability after freeze-thawing. PGCs obtained from the germinal gonade of 5.5~6 day (stage 28) chick embryos, using the MACS method were suspended in a freezing medium containing a freezing and protecting agents. Gonads were harvested from stage 28 chick embryos and pooled in groups of 10E embryos, contributing gonads to the cell suspension. The gonadal cells, including PGCs, were then frozen in 1 of the following cryoprotectant treatments: 2.5% EG, 5% EG, 10% EG, 2.5% PG, 5% PG, 10% PG, and 0% cryoprotectant as a control. Effects of exposure to vitrification solution and vitrification, with different concentrations of the cryoprotectant solution, were examined. After freezing and thawing, survival rates of the frozen-thawed PGCs from the 0, 2.5, 5, 10 and 15% EG plus FBS treatment were 44.24%, 64.51%, 85.63%, 80.51% and 73.52% (p<0.05), respectively. The viability of PGCs after freeze-thawing was significantly higher for 10% EG plus FBS treatment than for 10% PG + FBS treatment (p<0.05)(85.63% vs 66.81%). Therefore, these systems may contribute in the improvement of cryopreservation for a scarce species in birds preservation. This study established a method for preserving chicken PGC that enables systematic storage and labeling of cryopreserved PGCs in liquid N at a germplasm repository and ease of entry into a database. In the future, the importance for this new technology is that poultry lines can be conserved while work is being conducted on improving the production of germline chimeras.

• Journal of Aquaculture
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• v.13 no.3
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• pp.215-222
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• 2000

The primordial germ cell of the sweet fish was recognized from the 2-day old fry (tl : 0.66 cm), when it began to protrude into peritoneal cavity between meaonephric duct and gut. The primordial gonad, with the formation of genital ridge, developed on the 30-day old fry. Ovarian differentiation was identified by the presence of ovarian cavity and meiotic oocytes from the 90-day old fry (tl : 3.42 cm). Testicular differentiation was identified by the presence of spermatogonial cells with efferent duct from the 100-day old fry (11 : 4.50 cm). Hence the sweet fish belongs to the differentiated type of gonochoristic teleost.