• Reproductive and Developmental Biology
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• 제34권3호
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• pp.207-211
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• 2010

Current developments in IVF and animal cloning have resulted in increasing demand for large quantities of oocytes and ovarian follicles at specific stages of development. These medical and scientific needs may be met by developing an optimal culture system for preantral follicles. In this study, we investigated the growth of porcine preantral follicle cultures in different media and in the presence and absence of serum. Follicles were manually dissected from ovaries obtained from prepubertal gilts at a local slaughterhouse, and cultured for 3 days in M199 or NCSU23 medium supplemented with porcine FSH, transferrin, L-ascorbic acid and insulin. Follicle diameters were measured on day 1 and 3 of culture. In Experiment 1, the effect of supplementing culture medium with fetal calf serum (FCS) on porcine preantral follicle growth was examined. In the group of cultures supplemented with FCS, follicle diameter after 3 days of culture, survival rate and antrum formation rate in the FCS group were significantly higher than those of the control group. In Experiment 2, the effects of culture medium (M199 and NCSU23) on follicle growth were compared. Follicle diameters were increased in the M199 group, compared with those in NCSU23 (p<0.05), but we observed no significant differences in survival and antrum formation rates between cultures grown in the two media. In conclusion, supplementation of the culture medium with serum enhances preantral follicle growth and antrum formation, and M199 is superior to NUSU23 for porcine preantral follicle culture in vitro.

• Asian-Australasian Journal of Animal Sciences
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• 제22권1호
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• pp.35-41
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• 2009

This study was undertaken to evaluate how ${\beta}$-mercaptoethanol (bME), an exogenous antioxidant, interacts with preantral follicles cultured in vitro. Mouse primary or secondary follicles were cultured in glutathione (GSH)-free or GSH-containing medium supplemented with bME of various concentrations, and the growth of preantral follicles, the maturation of intrafollicular oocytes and preimplantation development after parthenogenesis were monitored. In experiment 1, 0, 25, 50 or 100 ${\mu}M$ bME was added to culture medium supplemented with 100 ${\mu}M$ GSH or not. When secondary follicles were cultured in GSH-free medium, no significant change in follicle growth was detected after bME addition. However, exposure to bME in the presence of GSH significantly inhibited both follicle growth and oocyte maturation. Such detrimental effect became prominent in primary follicles and bME strongly inhibited follicle growth in the absence of GSH. In conclusion, there are stage-dependent effects of bME on follicle growth and oocyte maturation, and selective use of antioxidants contributes to establishing an efficient follicle culture system.

• 한국발생생물학회지:발생과생식
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• 제20권3호
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• pp.227-235
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• 2016

Ultrastructural studies on oocyte development and vitellogenesis in oocytes, and the functions of follicle cells during oogenesis and oocyte degeneration were investigated to clarifyb the reproductive mechanism on vitellogenesis of Scapharca subcrenata using electron microscope observations. In this study, vitellogenesis during oogenesis in the oocytes occured by way of autosynthesis and heterosynthesis. Of two processes of vitellogenesis during oogenesis, the process of endogenous autosynthesis involved the combined activity of the Golgi complex, mitochondria and rough endoplasmic reticulum. However, the process of exogenous heterosynthesis involved endocytotic incorporation of extraovarian precursors at the basal region of the oolema of the early vitellogenic oocytes before the formation of the vitelline coat. In this study, follicle cells, which attached to the previtellogenic and vitellogenic oocytes, were easily found. In particular, the follicle cells were involved in the development of previtellogenic oocytes by the supply of nutrients, and vitellogenesis in the early and late vitellogenic oocytes by endocytosis of yolk precursors. Based on observations of follicle cells attached to degenerating oocytes after spawning, follicles of this species are involved in lysosomal induction of oocyte degeneration for the resorption phagosomes (phagolysosomes) in the cytoplasm for nutrient storage, as seen in other bivalves. In this study, the functions of follicle cells can accumulate reserves of lipid granules and glycogen particles for vitellogenesis from degenerating oocytes after spawning.

• 대한의생명과학회지
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• 제12권2호
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• pp.105-112
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• 2006

This study was undertaken to investigate the morphological changes between normal and atretic follicle after gamma irradiation and treatment of follicle stimulating hormone (FSH). The ovaries of each group of treated immature mice were prepared the paraffin sections after 0, 6, 12, and 24 hours (hrs) of those treatment. Hematoxylin-eosin (HE) stain, reticulin stain, and terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) immunohistochemical stain were performed on the each paraffin sections. As the results of HE staining, the condensed nuclei of oocytes were observed in the atretic primordial follicles, on the other hand the condensations of granulosa cell nuclei were prominent in the atretic primary, preantral, and antral follicles. Only the granulosa cells of atretic follicle were stained specifically with TUNEL staining but not stained in the theca cells, which suggested granulosa cells degenerated through apoptosis. In the reticulin staining, the basement membranes of atretic follicle which was stained weakly showed irregular structure and detachment from the follicles. The ratio of normal to atretic follicle in control and FSH treated group was about 33% but this ratio increased rapidly over 90% in the 6, 12, and 24 hrs group after the irradiation. It could be suggested that the gamma irradiation is the useful tool far the induction of follicle atresia and immunohistochemical staining methods are essential in the study of follicle atresia.

• 한국수정란이식학회지
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• 제27권3호
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• pp.141-147
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• 2012

This study was operated to establish induction using ultrasonography by estimating the relation of follicle size and estrus manifestation. Clinical estrus symptoms were observed 97.4% in cows and 87.5% in heifers when overall 55 cows were induced to estrus in a single dose of $PGF_{2{\alpha}}$ after verifying CL through ultrasonography, which means estrus hours among those 52 cows showing the clinical estrus symptoms were estimated 2.39 days on cows and for 2.37 days on heifers which showed no differences (p>0.05). The estrus manifestation hours according to the follicle size in cows didn't have any significance each other (p>0.05), though estrus hours was 54 hours (the shortest) with follicle size bigger than 10 mm and were made up within 69 hours. The estrus manifestation hours according to the follicle size in heifers didn't have any significance each other (p>0.05) and took around 42 hours (the shortest) with follicle size of 5mm (the smallest) and were made up within 66 hours. Follicles after $PGF_{2{\alpha}}$ injection were ovulated and assigned to many phases as follows; Group 1 (growing phase) - continuously growing into ovulation, Group 2 (growing and static phase) - delaying in growth after the growth of follicles, Group 3 (static and growing phase) - growing after growth delay, Group 4 (regressing and new growing phase) - the follicle is closed and a new follicle grows. In addition, the process of follicle development and estrus hours had no significance each other (p>0.05), though estrus manifestation hours in Group 1 and 2 was relatively short, and in Group 3 and 4 for a relatively long time. In the result of all above, the estrus manifestation hours after $PGF_{2{\alpha}}$ injection has no differences accoring to the follicle size in cows and heifers. Therefore, High pregnancy rate is obtained when practicing artificial insemination within 3 days in estrus or TAI in 72 to 80 hours after adminitrating $PGF_{2{\alpha}}$.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제36권1호
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• pp.7-15
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• 2010

Complex human tissues harbor stem cells and precursor cells, which are responsible for tissue development or repair. Recently, dental tissues such as dental pulp, periodontal ligament (PDL), dental follicle have been identified as easily accessible sources of undifferentiated cells. These tissues contain mesenchymal stem cells that can be differentiate into bone, cartilage, fat or muscle by exposing them to specific growth conditions. In this study, the authors procured the stem cell from pulp, PDL, and dental follicle and differentiate them into osteoblast and examine the bone induction capacity. Dental pulp stem cell (DPSC), periodontal ligament stem cell (PDLSC), and dental follicle precursor cell (DFPC) were obtained from human 3rd molar and cultured. Each cell was analyzed for presence of stem cell by fluorescence activated cell sorter (FACs) against CD44, CD105 and CD34, CD45. Each stem cell was cultured, expanded and grown in an osteogenic culture medium to allow formation of a layer of extracellular bone matrix. Osteogenic pathway was checked by alizarin red staining, alkaline phosphatase (ALP) activity test and RT-PCR for ALP and osteocalcin (OCN) gene expression. According to results from FACs, mesenchymal stem cell existed in pulp, PDL, and dental follicle. As culturing with bone differentiation medium, stem cells were differentiated to osteoblast like cell. Compare with stem cell from pulp, PDL and dental follicle-originated stem cell has more osteogenic effect and it was assumed that the character of donor cell was able to affect on differential potency of stem cell. From this article, we are able to verify the pulp, PDL, and dental follicle from extracted tooth, and these can be a source of osteoblast and stem cell for tissue engineering.

• Applied Microscopy
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• 제24권4호
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• pp.52-60
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• 1994

The developing ovarian oocyte of Dendrolimus spectabilis has been studied by using electron microscopical techniques. After yolk formation the vitelline membrane was laid down in the intercellular space between the follicle cell and the oocyte. But before the vitelline membrane formation the granules with high electron density that the vitelline membrane precusor are observed in the follicle cell. At the late vitellogenesis stage these granules were transported into the intercellular space between the follicle cells and the oocyte. These granules fuse to each other and larger bodies which eventually produce the vitelline membrane. The vitelline membrane was distinguished into the light inner and dark outer membrane. Next the chorion was laid down. It was apparent that the chorion was laid down in the intercellular space immediately adjacent to the vitelline membrane, and that it was formed by the follicle cells only.

• 한국발생생물학회지:발생과생식
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• 제4권1호
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• pp.13-17
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• 2000

FSH는 미성숙 설치류의 난포성장을 촉진하며, 강소형성 난포의 퇴화비율을 감소시킨다. 본 연구는 미성숙 생쥐에 난포성숙호르몬을 투여한 후 유발되는 난포의 조직학적인 변화를 규명하기 위해 시행되었다. 3주령의 ICR생쥐에 10 i.u.의 재조합 난포자극호르몬을 복강주사한 후 1일, 2일, 3일에 좌측 난소의 조직학적 변화를 관찰하였다. 강소형성전 난포의경우 FSH처리 후 시간에 따라 퇴화난포의 비율이 증가하였으나 강소형성 난포의 경우에는 유의한 변화를 보이지 않았다. 퇴화되는 양상은 난포 내 세포자연사하는 과립세포의 증가, 대식세포 및 다형다핵백혈구의 증가 등이 관찰되었다. 이상의 결과로 보아, 과량의 FSH처리 후에 유발되는 난포의 퇴화는 과립세포의 세포자연사뿐 아니라 급성 염증반응을 수반하는 것으로 생각된다.

• 환경생물
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• 제33권4호
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• pp.419-424
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• 2015

두꺼비 (Bufo gargarizans)의 난소주기를 파악하기 위해 암컷 성체를 대상으로 gonadosomatic index (GSI)와 난소 내 여포난자의 크기와 난황 축적 정도를 기준으로 발달과정을 연중 조사하였다. 4월에는 난소 무게와 GSI가 가장 낮게 나타났으며 모든 여포난자들은 난황 축적 전단계의 상태로 존재하여 난황 형성이 중단된 것으로 판단된다. 난소 무게와 GSI가 증가하기 시작한 5월의 난소에서는 난황 축적 전기단계의 여포난자가 출현하였고 6월에는 난황 축적 중기단계와 난황 축적 전단계의 여포난자가 존재하여 난소 무게와 GSI도 증가하여 나타났다. 이러한 현상은 이 시기에 난황 축적현상이 진행되는 것을 의미하며 난소 무게와 GSI가 급격히 증가하기 시작한 9월에는 난황 축적 중기단계의 여포난자들과 난황 축적이 거의 완성된 난황 축적 후기단계의 여포난자들이 존재하였다. 난황 축적을 마치고 성장이 완료된 여포난자들은 10월부터 일부 출현하기 시작하여 동면기인 12월에 급격히 증가하였으며 2월의 난소에서는 성장이 완료된 여포난자가 전체적으로 존재하여 여포난자의 성장기에는 난소 내의 모든 여포난자들이 동시적 (synchronized)으로 진행되지 않고 각각의 여포난자에 따라 진행되다가 배란시기에 성장이 완료된 상태를 유지하는 난소주기를 나타내었다.

• 한국환경생태학회지
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• 제28권3호
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• pp.287-292
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• 2014

한국산개구리(Rana coreana)의 난소주기를 파악하기 위해 암컷 성체를 대상으로 gonadosomatic index(GSI)와 난소 내 여포난자의 크기와 난황축적 정도를 기준으로 발달과정을 연중 조사하였다. 난소무게와 GSI는 3월부터 5월까지 가장 낮게 나타났으며 모든 여포난자들은 난황축적 전단계의 상태로 존재하여 난황형성이 중단된 것으로 판단된다. 난소무게와 GSI가 증가하기 시작한 6월의 난소에서는 난황축적 전기단계의 여포난자가 출현하였고 8월에는 난황축적 중기단계와 난황축적 전단계의 여포난자가 존재하여 난소무게와 GSI도 증가하여 나타났다. 이러한 현상은 이 시기에 난황축적 현상이 활발하게 진행되는 것을 의미하며 난소무게와 GSI가 높게 나타난 9월에서 11월까지는 난황축적 중기단계의 여포난자들과 난황축적이 거의 완성된 난황축적 후기단계의 여포난자들이 존재하였다. 동면중인 12월부터 난황축적을 마치고 성장이 완료된 여포난자들이 출현하였으며 2월의 난소에서는 성장이 완료된 여포난자가 전체적으로 존재하여 여포난자의 성장기에는 난소내의 모든 여포난자들이 동시적(synchronized)으로 진행되지 않고 각각의 여포난자에 따라 진행되다가 배란시기에 성장이 완료된 상태를 유지하는 난소주기를 나타내었다.