• Journal of Ginseng Research
• /
• v.19 no.2
• /
• pp.171-174
• /
• 1995

Freshly harvested American ginseng (Panax quinquefolium L.) seeds were stratified at two locations over each of three years. Seed development and temperature in the stratification boxes were investigated until the seed was removed 12 months later and direct-seeded in the field. During stratification and seeding (14 months) three embryo stages were identified. In Stage I of 250 days (Sept. to mid-May) embryo length increased from about 0.5 to 1.0 mm: in Stage II of 100 days (mid-May to late Aug. when seeded) length increased to 2.0 mm and in Stage III (late Aug. to late Nov.) length increased to 5.3 mm. Excerpt split width could also be placed in three stages. Changes in embryo length correlated with embryo endosperm length ratio. Insertion compression tests showed that the excerpt softened rapidly in late Stage II and throughout Stage III. The stratification box temperatures at all depths (10, 25 and 50 cm) never exceeded -2$^{\circ}C$ even when the air temperatures dropped to -13$^{\circ}C$ and were, therefore not damaging to the seeds.

• Proceedings of the KSAR Conference
• /
• 2003.06a
• /
• pp.69-69
• /
• 2003

Although effect of TGF$\beta$$_1$ on preimplantation embryo development was reported at mice, little information relevant to this subject is known in bovine. The objectives of this study were to investigate TGF$\beta$$_1$, and TGF$\beta$$_1$ receptors type I and II expression, known as important factors in the embryo development, at unfertilized oocytes and fertilized embryos that will be used as basic data to be compared to NT embryos. We postulated that TGF$\beta$$_1$ may have a beneficial effect on the preimplantation embryo and show different expression patterns as embryo stages change. We have used immunocytochemistry to investigate the presence in unfertilized oocytes and preimplantation embryos of TGF$\beta$$_1$ and the essential components of the TGF$\beta$$_1$ signalling pathway, TGF$\beta$$_1$ receptors type I and II. We found that both receptors, as well as TGF$\beta$$_1$, were present in the unfertilized oocytes. This indicates that TGF$\beta$$_1$, is a maternally expressed protein. At the morulae and blastocyst stages the TGF$\beta$$_1$ receptor type II was not present, but the TGF$\beta$$_1$ receptor type I was present at both stages and we can confirm the TGF$\beta$$_1$ expression of high level at 8-cell stage. These findings support our hypothesis that the TGF$\beta$$_1$, and TGF$\beta$$_1$ receptors may interact with the oocyte and preimplantation embryo, and that TGF$\beta$$_1$ signalling may be important for the development of the oocyte and the preimplahtation embryo.

• Reproductive and Developmental Biology
• /
• v.30 no.2
• /
• pp.75-79
• /
• 2006

The developmental regulation of the preimplantation mammalian embryos is a fundamental step for preparing the implantation and it may be regulated by several autocrine and paracrine factors including platelet-activating factor. PAF improved the embryonic survival and implantation but its role during blastocyst development is still largely unknown. In this study, the effects and the possible pathway of PAF on developmental regulation of blastocyst to hatching stage were investigated. Developmental pattern in hatching embryo was a concentration-response curve showing maximal activity at 1 nM PAF, with decreasing activity at higher concentrations. $50{\mu}M$ 1-(5-isoquinolimnesulfonyl)-2-methylpiperazinme dihydrochloride (H-7), a PKC inhibitor, inhibited the progression of blastocyst to hatching embryo. In addition H-7 blocked the PAF effects on the blastocyst development. Besides tetradecanoylphorbol acetate (TPA), a PKC activator stimulated development of blastocyst to the hatching stage. These finding revealed that PAF support the blastocyst development to the hatching embryo. Also it is suggested that PAF action pathways in hatching supporting include the PKC signaling pathway.

• Korean Journal of Animal Reproduction
• /
• v.22 no.1
• /
• pp.101-104
• /
• 1998

This study examined the effects of growth factors in TCM199 on bovine 1-cell embryos development in vitro. After 6 day to 11 day in culture, 15.8%(19/120), 15.3%(20/130), 21.8%(35/160), 27.0%(56/207), 26.3%(53/201) and 30.7%(40/130) of the 1-cell embryos developed into expanding blastocysts in su, pp.ementing TCM199 with control, insulin, IGF-I, IGF-II, FGF and EGF, respectively. Hatching rate of 1-cell embryos in su, pp.ementing TCM199 with FGF, EGF and IGF-II were 21.4%(53/247), 20.3%(42/206) and 16.8%(41/243), respectively. The beneficial effect of growth factors on embryo development in vitro could be duplicated. These data indicate that the presence of FGF, EGF or IGF-II in the culture medium is beneficial for embryo development in vitro and accelerate cell differentiation.

• Journal of Embryo Transfer
• /
• v.9 no.2
• /
• pp.167-172
• /
• 1994

This experiment was arried out to investigate the development of ea4y rabbit embryos in vivo. Twenty-six New Zealand White does were superovulated by treatment with PMSG(Intervet Co; I. M single injection, 150. U./rabbit) followed 3 day later by simultaneous I.V injection of 100 I.U HCG (Intervet Co, )and natural service with fertile male. All of does was killed at the specific times (24, 27, 30, 36, 42, 50 and 93 h post-hCG) to find out the early embryonic development in vivo respectively. Embryos at the specified stages of development were obtained at the following times after injection of hCG; one-ceH at 24 h, two-cell at 24~27h, four-cell at 27~36 h, morulae at 50 h and early blasto-cyst at 93 h and expanded or hatching blastocyst at 144 h. Number of embryos recovered per rabbit superovulated was 26.1 and average of recovery rate was 83.7%. The results suggest that superovulation was efficient for the increase of embryo number in rabbits, and as shown in results, asynchronous cleavage was prevalent among the recovered embryos.

• Horticultural Science & Technology
• /
• v.31 no.4
• /
• pp.483-489
• /
• 2013

The abnormal seedlings, a common physiological anomalies, emerged during embryo rescue severely restricted grape breeding. To enhance the efficiency of the seedless grape breeding by reducing the production of abnormal seedlings in the course of embryo rescue, we investigated the effects of genotype, media type, embryo style, pre-chilling on the deformity rate of the abnormal seedlings during embryo rescue. The abnormal seedlings were firstly classified into seven categories based on their morphology. Our results indicated that the emergence of abnormal seedlings was highly dependent on the female parent genotype. Polyembryony was advantageous to diminish the number of abnormal plantlets and the germination rate of embryo was 100%. We also found that pre-chilling treatment could reduce the number of abnormal plantlets and promote the embryo germination. The abnormal plantlets were reduced significantly by the addition of $ZnSO_4$ $10{\mu}mol{\cdot}L^{-1}$ or mashed-banana $500mg{\cdot}L^{-1}$ to either embryo development or germination media. Transferring the abnormal seedlings onto the suitable fresh media in 4 weeks after embryo germination provided an effective way to transform them into normal seedlings.

• Korean Journal of Animal Reproduction
• /
• v.13 no.3
• /
• pp.141-148
• /
• 1989

The effects of seeding method and optimum time for freezing embryos according to the developmental stages on embryo survival rates after rapid freezing were determined using the FDA-test. The summarized results are as follows : 1. In the rapid freezing of embryos, the sucrose added medium together with Co-seeding or non-seeding showed the FDA scores of 4.67 and 4.20, respectively, but, raffinose addition obtained FDA scores of 4.27 and 3.97. 2. The developmental stage of embryos at freezing was most critical on the survival of embryos after thawing. Higher FDA scores were obtained in the order of blastocyst stage(4.94), morula stage(3.82) and ealy stage(2.65) in sucrose added medium. The same trend was observed in the raffinose added medium with an order or 4.91, 4.47 and 2.32. 3. Microscopic study of embryo before freezing and post-thawing indicated that the embryo showed shrinkage within 5 minutes after the embryo was transfer to the freezing medium. When thawed embryo was tranfered to the dilution medium, swelling of the embryo was observed and there after it reshrank indicating the removal of cryoprotectant from the embryo. The size of the embryo recovered to the original state when it was moved into a PBS-solution.

• Clinical and Experimental Reproductive Medicine
• /
• v.44 no.1
• /
• pp.40-46
• /
• 2017

Objective: To describe in vitro development of human embryos derived from an individual with a homozygous pathogenic variant in NLRP7 (19q13.42) and recurrent hydatidiform mole (HM), an autosomal recessive condition thought to occur secondary to an oocyte defect. Methods: A patient with five consecutive HM pregnancies was genomically evaluated via next generation sequencing followed by controlled ovarian hyperstimulation, in vitro fertilization (IVF) with intracytoplasmic sperm injection, embryo culture, and preimplantation genetic screening. Findings in NLRP7 were recorded and embryo culture and biopsy data were tabulated as a function of parental origin for any identified ploidy error. Results: The patient was found to have a pathogenic variant in NLRP7 (c.2810+2T>G) in a homozygous state. Fifteen oocytes were retrieved and 10 embryos were available after fertilization via intracytoplasmic sperm injection. Developmental arrest was noted for all 10 embryos after 144 hours in culture, thus no transfer was possible. These non-viable embryos were evaluated by karyomapping and all were diploid biparental; two were euploid and eight had various aneuploidies all of maternal origin. Conclusion: This is the first report of early human embryo development from a patient with any NLRP7 mutation. The pathogenic variant identified here resulted in global developmental arrest at or before blastocyst stage. Standard IVF should therefore be discouraged for such patients, who instead need to consider oocyte (or embryo) donation with IVF as preferred clinical methods to treat infertility.

• Development and Reproduction
• /
• v.21 no.2
• /
• pp.205-213
• /
• 2017

The aim of this study was to determine the effect of additional alpha-linolenic acid (ALA) supplementation during in vitro maturation (IVM) and culture (IVC) on nucleic maturation and embryo development of pigs. Cumulus-oocyte complexes (COCs) were incubated in IVM medium containing different concentration of ALA (25, 50 and $100{\mu}M$) for 44 h. After in vitro maturation, nuclear maturation of oocytes were evaluated by aceto-orcein stain. Mature oocytes with $50{\mu}M$ ALA were fertilized and cultured in IVC medium with ALA (25, 50 and $100{\mu}M$) during early-embryogenesis (48 hours after fertilization). Then, embryos were cultured with $25{\mu}M$ ALA during early embryogenesis and/or late embryogenesis (120 hours after early-embryogenesis). In results, oocyte maturation were significantly increased by $50{\mu}M$ ALA treatment groups compared with control groups (p<0.05). Treatment of $25{\mu}M$ ALA during early-embryogenesis enhanced cleavage rate of embryo compared with other groups (p<0.05), whereas formation and total cell number of blastocyst had no significant difference. Similarly, cleavage rate of embryos were increased by $25{\mu}M$ ALA supplement during early- or late-embryogenesis than ALA treatment both stage of embryogenesis (p<0.05), but did not influence to blastocyst formation. Interestingly, total cell number of blastocyst were enhanced in ALA treatment group during early-embryogenesis. These findings indicated that ALA supplement enhance the nuclear maturation of oocyte and embryo development, however, excessive ALA could negatively influence. Therefore, we suggest that ALA is used for improvement of in vitro production of mammalian embryo and further study regarding with functional mechanism of ALA is needed.

• Proceedings of the Botanical Society of Korea Conference
• /
• 1999.07a
• /
• pp.85-89
• /
• 1999

Somatic embryogendesis is one of good examples of the basic research for plant embryo development as well as an important technique for plant biotechnology. This paper describes the direct somatic embryogenesis from zygotic embryos of Panax ginseng is reversely related to normal axis growth of zygotic embryos by the experiment of various chemical treatments. Under the normal growth condition, the apical tips of embryo axis produced an agar-diffusible substance, which suppressed somatic embryo development from cotyledons. Although the cells of zygotic embryos were released from the restraint of embryo axis, various factors were still involved for somatic embryo development. Electron microscopic observation revealed that the ultrastructure of cells of cotyledon epidermis markedly changed before initiation of embryonic cell division, probably indicating reprogramming events into the cells embryogenically determined state. Polar accumulation of endogenous auxin or cell-cell isolation by plasmolysis pre-treatment is the strong inducer for the somatic embryo development. The cells for the process of somatic embryogenesis might be determined by the physiological conditions fo explants and medium compositions. Direct somatic embryos from cotyledons fo ginseng were originated eithrer from single or multiple cells. The different cellular origin of somatic embryos was originated either from single or multiple cell. The different cellular origin of somatic embryos was depended on various developmental stages of cotyledons. Immature meristematic cotyledons produced multiple cell-derived somatic embryos, which developed into multiple embryos. While fully mature cotyledons produced single cell-derived single embryos with independent state. Plasmolysis pretreatment of cotyledons strongly enhanced single cell-derived somatic embryogenesis. Single embryos were converted into normal plantlets with shoot and roots, while multiple embryos were converted into only multiple shoots. GA3 or a chilling treatment was prerequisite for germination and plant conversion. Low concentration of ammonium ion in medium was necessary for balanced growth of root and shoot of plantlets. Therefore, using above procedures, successful plant regeneration of ginseng was accomplished through direct single embryogenesis, which makes it possible to produce genetically transformed ginseng efficently.