• 한국수정란이식학회지
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• 제27권4호
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• pp.193-203
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• 2012

The development of embryos reconstructed by nuclear transfer is dependent upon numerous factors including the type of recipient cell, method of enucleation, the type of donor cell, method of reconstruction, activation, the cell cycle stage of both the donor nucleus and the recipient cytoplasm and the method of culture of the reconstructed embryos. Many of these points which have been reviewed extensively elsewhere (Sun and Moor, 1995; Colman, 1999; Oback and Wells, 2002; Renard et al., 2002; Galli et al., 2003b), here we will concentrate on main area, the production of suitable cytoplast and nuclear donor, nuclear-cytoplasmic coordination, oocyte activation, culture of reconstructed embryos, and the effects that this may have on development.

• Clinical and Experimental Reproductive Medicine
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• 제21권3호
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• pp.325-330
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• 1994

The study has been carried out in order to evaluate the effects of embryonic stage, and cryopreservation method on the rates of viability and development of the cryopreserved mouse early embryos. The results were as following:In the treatment steps of cryoprotectant, for the fertilized oocyte with pronucleus(PN), 2-step was better than the others. And for the other embryos, 4-step was better than 2- or 3-step. In respect to the embryonic stage, as the embryos developed from fertilized oocytes to 8-cell embryos, the rates of viability and development were increased higher. Therefore, 8-cell embryo was better stage than the others. In respect to the kind of cryoprotectants, PROH was better than DMSO for the fertilized oocyte, as a cryoprotectant. DMSO, for the 2-cell embryos and PROH and DMSO for the 4- and 8-cell embryos were suitable for cryopreservation.

• 한국발생생물학회지:발생과생식
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• 제25권3호
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• pp.173-183
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• 2021

The incidence of infertility among individuals of reproductive age has been growing due to genetic and environmental factors, and considerable research efforts are focused on solving this issue. Ovarian development is an overly complex process in the body, involving the interaction between primordial germ cells and gonad somatic cells. However, follicles located in the center of the in vitro ovary are poorly formed owing to ovarian complexity, nutrient deficiency, and signaling deficiency. In the present study, we optimized methods for dissociating gonads and culture conditions for the in vitro generation of miniaturized ovaries. The gonads from embryos were dissociated into cell masses and cultured on a Transwell-COL membrane for 3-5 weeks. Approximately 12 follicles were present per in vitro ovary. We observed that miniaturized ovaries successfully matured to MII oocytes in vitro from 150 to 100 ㎛ gonad masses. This method will be useful for investigating follicle development and oocyte production.

• Reproductive and Developmental Biology
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• 제29권2호
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• pp.117-120
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• 2005

This study was conducted to establish the optimal temperature condition before oocyte activation in B6m F1 mouse. In experiment 1, two embryo culture media (CZB vs KSOM) were evaluated for the development of activated mouse oocytes. Parthenogenetic embryos cultured in KSOM showed better blastocyst development than ones cultured in CZB $(56.2\%\;vs\;81.0\%\;p<0.01)$. Two-hour of pre-incubation before activation significantly reduced the number of hatched blastocysts in KSOM $(22.0\%\;versus\;8.8\%\;p<0.05)$. In experiment 2, recovered oocytes were pre-incubated at different temperature conditions before activation. The experimental groups were divided by 5 as follows. Group A: pre-incubation for 120 min at $37^{\circ}C$, Group B: pre-incubation at $37^{\circ}C$ for 90 min then at $25^{\circ}C$ for 30 min, Group C: pre-incubation at $37^{\circ}C$ for 60 min then at $25^{\circ}C$for 60 min, Group D: pre-incubation at $37^{\circ}C$ for 30 min then at $25^{\circ}C$ for 90 min, and Group E: pre-incubation at $25^{\circ}C$ for 120 min before activation. Group A $(67.6\%)$ and B $(66.7\%)$ showed better development to the blastocyst stage than other groups $(Group\;C:\;50.0\%\;Group \;D:\;49.2\%\;Group\;E:\;33.3\%,\;p<0.05)$. The present study indicates that the temperature before activation affects the development of B6D2 F1 mouse parthenogenetic oocytes and exposure to room temperature should be limited to 30-min when the oocytes are left in HEPES-buffered medium for micromanipulation.

• Asian-Australasian Journal of Animal Sciences
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• 제29권8호
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• pp.1065-1074
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• 2016

Growth factors play an important role during early ovarian development and folliculogenesis, since they regulate the migration of germ cells to the gonadal ridge. They also act on follicle recruitment, proliferation/atresia of granulosa cells and theca, steroidogenesis, oocyte maturation, ovulation and luteinization. Among the growth factors, the growth differentiation factor 9 (GDF9) and the bone morphogenetic protein 15 (BMP15), belong to the transforming growth factor beta (TGF-${\beta}$) superfamily, have been implicated as essential for follicular development. The GDF9 and BMP15 participate in the evolution of the primordial follicle to primary follicle and play an important role in the later stages of follicular development and maturation, increasing the steroidogenic acute regulatory protein expression, plasminogen activator and luteinizing hormone receptor (LHR). These factors are also involved in the interconnections between the oocyte and surrounding cumulus cells, where they regulate absorption of amino acids, glycolysis and biosynthesis of cholesterol cumulus cells. Even though the mode of action has not been fully established, in vitro observations indicate that the factors GDF9 and BMP15 stimulate the growth of ovarian follicles and proliferation of cumulus cells through the induction of mitosis in cells and granulosa and theca expression of genes linked to follicular maturation. Thus, seeking greater understanding of the action of these growth factors on the development of oocytes, the role of GDF9 and BMP15 in ovarian function is summarized in this brief review.

• 한국동물생명공학회지
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• 제35권2호
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• pp.119-141
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• 2020

The Dromedary camel (Camelus dromedaries) is an important species because of its ability to produce good quality meat, milk, and fibers under harsh environmental conditions. Camels are also crucial for transportation, racing, and as draft animals in agriculture. Therefore, dromedary camels play a critical role in the economy for millions of people living in the arid part of the world. The inherent capability of camels to produce meat and milk is highly correlated with their reproductive performance. Compared with other domestic species, the reproductive efficiency in camelids is low. Although recent reproductive technologies such as in vitro fertilization (IVF) and somatic cell nuclear transfer (SCNT) have been successfully applied to camelids and the birth of live offspring following these technologies has been reported; in vitro embryo production (IVP) has lagged in this species. The development of the IVP system for dromedary camels may be a useful tool for the genetic improvement of this species. IVP in farm animals includes three main steps; in vitro maturation (IVM) of an oocyte, IVF of a matured oocyte, and in vitro culture (IVC) of fertilized oocyte up to the blastocyst stage. This review aims to summarize various factors that influence oocyte quality, IVM, and in vitro embryo development in dromedary camel.

• Clinical and Experimental Reproductive Medicine
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• 제26권3호
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• pp.369-375
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• 1999

Objective: To evaluate the effect of serum obtained before and after treatment for endometriosis on in vitro fertilization and development of two cell mouse embryo. Design: Pretreatment and posttreatment comparoson of fertilization of mouse oocyte and embryo development in serum supplement from patients with endometriosis; result were compared using Stuent T-test analysis. Method: Infertility Clinic, Department of Obstetrics and Gynecology, Collage of Medicine, Won kwang university, Korea. Patients was chosed eleven consecutive women with endometriosis. Interventions was all patient underwent laparoscopic or conservative surgery. This was followed by a 6-month course of burserelin acetate $900{\mu}g/d$. Main outcome was measured total number of fertilization and embryo that was fertilization after 24 hours and reached blastocyst stage after 72 hours of incubation were compared before and after treatment. Result: Before treatment, 47% of the oocyte were fertilized and 31% of the embryo reached blastocyst stage. After treatment, Significantly more fertilized and Significantly more embryo developed to blastocyst on the stage I and II of endometriosis. Conclusion: The fertilization and embryo toxicity of serum samples from patients with endometriosis is lost after treatment.

• 한국패류학회지
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• 제26권2호
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• pp.177-184
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• 2010

The structure of the ovary, ultrastructure of oocytes and morphological characteristics of vitellogenesis during oogenesis in female Gomphina veneriformis were investigated in clams collected from coastal waters of Samchok, Gangwon-do, Kore. In the previtellogenic oocytes, the Golgi complex was involved in the formation of a number of vacuoles. In the early vitellogenic oocytes, lipid droplets appeared among the Golgi complex, endoplasmic reticulum, and mitochondria in the cytoplasm of the oocyte were involved in the formation of lipid droplets. Coated vesicles, resulting from endocytosis appeared at the basal region of the early vitellogenic oocyte. The uptake of nutritive materials in the coated vesicles formed by receptor-mediated endocytosis appeared through the formation of coated endocytotic pits on the oolemma. In the late vitellogenic oocytes, large yolk granules were formed by a combination of small yolk granules. In the mature oocyte, a mature yolk granule in composed of three components: crystaline core, electron lucent cortex, and a limiting membrane. According to cytological and histological observations, vitellogenesis occurred by way of endogenous autosynthesis and exogenous heterosynthesis. Autosynthesis involved the conbined activities of the Golgi complex, mitochondria, rough endoplasmic reticulum, whereas heterosynthesis involved endocytotic incorporation of extraovarian precursors at the basal region of the early vitellogenic oocyte. The follicle cells which was attached to oocytes, were involved in the development of the previtellogenic and early vitellogenic oocytes as a kind of nutritive cells containing a number of glycogen particles and lipid droplets in the cytoplasm.

• Clinical and Experimental Reproductive Medicine
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• 제47권4호
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• pp.269-276
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• 2020

Objective: We investigated the impact of tyrosine kinase inhibitor (imatinib or dasatinib) coadministration with cyclophosphamide (Cp) on preantral follicle development in an in vitro mouse model. Methods: Seventy-three female BDF1 mice were allocated into four experimental groups: group A, saline; group B, Cp (25 mg/kg); group C, Cp (25 mg/kg) and imatinib (7.5 mg/kg); and group D, Cp (25 mg/kg) and dasatinib (7.5 mg/kg). Preantral follicles were isolated and cultured in vitro up to 12 days. Final oocyte acquisition and spindle integrity of metaphase II (MII) oocytes were assessed. Levels of 17β-estradiol and anti-Müllerian hormone (AMH) in the final spent media were measured by enzyme-linked immunosorbent assays, and the mRNA levels of Star, Sod1, Mapk3, and Casp3 in the final follicular cells were quantified by real-time polymerase chain reaction. Results: The percentage of MII oocytes per initiated follicle, the proportion of MII oocytes with normal spindles, and the 17β-estradiol level were similar in all four groups. The median AMH level in group B (7.74 ng/mL) was significantly lower than that in group A (10.84 ng/mL). However, the median AMH levels in group C (9.96 ng/mL) and group D (9.71 ng/mL) were similar to that in group A. The mRNA expression levels of Star, Sod1, Mapk3, and Casp3 were similar in all four groups. Conclusion: Coadministration of imatinib or dasatinib with Cp could preserve AMH production capacity in this in vitro mice preantral follicle culture model, and it did not affect MII oocyte acquisition.

• 한국어류학회지
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• 제19권1호
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• pp.16-23
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• 2007

한국산 버들치속 (Rhynchocypris) 어류인 버들치 (Rhynchocypris oxycephalus)와 금강모치 (Rhynchocypris kumgangensis) 난모세포의 난막구조에 대해 광학현미경과 전자현미경으로 조사하였다. 두 종에 있어서 난형성과정은 비슷했으나 난모세포를 둘러싸는 여포세포층(follicular layer)에 있어서는 차이를 보였다. 버들치는 난황포(yolk vesicle)시기에 있어 여포세포층은 안쪽에 입방형 또는 둥근모양의 세포층(inner follicular layer)이 난막위에 형성되고 그 바깥쪽으로 편평세포층(outer follicular layer)의 2층으로 이루어져 있었다. 난모세포의 발생이 진행됨에 따라 inner follicular layer의 입방형세포는 원주형세포(columnar cell)로 바뀌게 된다. 난황구(yolk granule)시기에 원주형세포는 세포질에 부착물질인 mucin을 분비해서 난세포 전체를 둘러싸게 된다. 반면에 금강모치의 경우 버들치와 마찬가지로 난황포시기에 안층의 입방형 또는 둥근모양의 세포층과 바깥층의 편평세포층을 가지게 되지만 안층의 세포는 더 이상 변화를 보이지 않았으며, 부착물질 또한 형성되지 않았다. 이처럼 한국산 버들치속에 있어 난막의 구조적 차이는 두 종간에 뚜렷한 분류형질로도 이용될 수 있을 뿐 아니라 그들의 서식처 및 산란습성과도 연관이 있는 것으로 생각된다.