• Proceedings of the Ginseng society Conference
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• 1974.09a
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• pp.9-22
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• 1974

Unlike the tissue culture in animals and human being, in higher plants various parts of the plant are cultured for varied purposes, and they are named variously depending on which parts are used as explants or what purposes they are cultured for. Followings are some of the names of culture used frequently: organ culture, tissue culture, callus culture, single cell culture, meristem culture, mericlone culture, ovary culture, ovule culture, embryo culture, endosperm culture, anther culture, pollen culture, protoplast culture, etc.. As the names of the culture indicate, in some kinds of culture the explants used for culture are actually not tissues, but organs, single cells, or protoplasts. It seems, however, convenient to call all of the above-mentioned cultures grossly as tissue culture. Several kinds of tissue culture were attempted using Panax ginseng as material and some of the results were summarized below. 1. Callus culture After dormancy of the sed was broken, whole embryo or parts (hypocotyl, cotyledon and epicotyl) of partly grown embryo were cultured in the media supplemented with growth regulators. Rapid swelling occurred in a few weeks, but most of the swelling was observed only in the basal part of epicotyl, changes in the other parts of embryo appearing in much later stages. The swelling or increase in size, however, was resulted not from the divisions of cells, but from the mere expansion of cell. Real calli were formed about two months after inoculation of explants. Callus tissues developed from cortex, pith, and vascular bundle in the cases of hypo- and epicotyl, from mesophyl tissue in the case of cotyledon. Shoots developed more easily from cotyledons regardless of whether they are detached from or attached to the embryo proper. 2. Culture in the Knudson C medium When cotyledons, detached from or attached to the embryo proper, were cultured in the growth regulator-free Knudson C medium comprision only several kinds of mineral compounds and sucrose, shoot primordium or callus developed profusely and finally plantlets were produced directly from shoot primordium or indirectly through callus. In this medium epidermal cells as well as mesophyl cells of the cotyledon became meristematic and divided, changing into multinucleate cells or multicellular bodies, developing eventually into either shoot primordia or calli. 3. Anther culture Anthers were cultured in the media supplemented with various growth regulators applied singly or in combinations. Callus was formed mostly in the connective tissue of anther. Cells of anther wall layers changed in appearance, but no division occurred. Microspores of all stages in development were not changed, ruling out the possibility that microspore-originated callus might be formed. 4. Isolation of protoplast Protoplasts were isolated from young root, leaf, and epicotyl, using 0.7M D-mannitols as osmoticum and using macerozyme and cellulase respectively for maceration and digestion of the cell wall. Production in large number of naked intact protoplast was rather difficult as compared with other plant species. Fusion of protoplasts occurred infrequently mainly due to the fewer number of naked protoplasts in the solution.

• Journal of Embryo Transfer
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• v.31 no.3
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• pp.249-254
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• 2016

It is imperious to know the reproductive parameters of the subsisting swine breed for profitable farming in Bangladesh. This study was designed to compare the reproductive parameters of sows at hilly areas between the local and crossbred at farm and backyard level. A total of 116 breedable sows were included. The data on age at puberty, litter size, birth weight, weaning weight, gestation length, interval between farrowing and number of piglets born per sow in local and crossbred sows were determined and direct questionnaire method was used for data collection. The present study revealed that age at puberty between farm and backyard system of crossbred sows was $8.88{\pm}0.153$ and $9.85{\pm}0.221$ months, respectively. The farrowing interval was shorter in crossbred sows which were kept in farming system rather than backyard system. The interval between farrowing and onset of estrus was shorter in crossbred sows. The gestation length was almost similar between backyard and farm condition. The number of litter size per sow per farrowing was $8.45{\pm}0.41$ in crossbred and $7.17{\pm}0.250$ in local pigs at backyard system. The birth weight and weaning weight were more in farm conditions than backyard system. The age at puberty in local breed was lower than the crossbred and litter size was $8.45{\pm}0.407$ in crossbred which was higher than local breed $7.17{\pm}0.250$ in backyard system. These results suggest that reproductive parameters of local sow need to be improved for better production and crossbred sows should be reared for obtaining desired productivity.

• Journal of Embryo Transfer
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• v.26 no.1
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• pp.1-7
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• 2011

The purpose of this study was to determine the relationship between conception rate and other parameters (body condition score; BCS, progesterone concentrations and follicle size) before estrus induction with CIDR(intravaginal progesterone-releasing controlled internal drug release). The conception rate in cows with < 2.75, 2.75 to 3.25, and 3.25 <, BCS regardless of AI (artificial insemination) time was 46.6%, 63.3%, and 46.6% at CIDR insertion, respectively. The conception rate regardless of BCS was 54.9% in cows inseminated based on detected estrus, and 48.7% in cows inseminated at 72 to 80 hours (timed artificial insemination, TAI) after removal of CIDR. The conception rate regardless of AI time was 40.0% in cows with low progesterone concentrations (less than 1.0 ng/ml), and 56.6% in cows with high progesterone concentrations (more than 1.0 ng/ml) at CIDR injection. The conception rate regardless of progesterone concentrations was 53.8% in cows inseminated based on detected estrus, and 38.0% in cows of TAI after removal of CIDR. The conception rate regardless of AI time was 43.3% in cows with small follicle (less than 5 mm), 53.3% in cows between 5 mm to 10 mm of follicle, and 63.3% in cows with large folliclc (more than 10 mm) at CIDR injection, respectively. The conception rate regardless of follicle size was 58.4% in cows inseminated based on detected estrus, and 45.9% in cows of TAI after removal of CIDR. These results indicated that if the cows with BCS 2.75 to 3.25, active corpus luteum, and/or large dominant follicle (more than 10 mm) are used for estrus induction, the conception rate will be greater.

• Journal of Embryo Transfer
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• v.22 no.3
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• pp.191-194
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• 2007

The purpose of the present study was to set up basic information of size and status of ovaries by using ultrasonography to retrieve in vivo matured oocytes with ovum pick- up method. Ovaries were collected from the abattoir in Jeju in May and June which is breeding season. When the size of ovaries on ultrasonography was compared with real size measured by caliper, no significant difference was shown (p<0.05). The number of preovulatory follicles (>21mm) was investigated with ultrasonography and naked eyes. Ultrasonography group had 0.83 preovulatory follicles per ovary and naked-eye group had 0.75 preovulatory follicle per ovary and their average size was 2.86 cm and 2.34 cm, respectively. The average number of follicle was 4.25 with ultrasonography and 4.38 with naked eyes. There was no significant difference considering the size of follicle and number of follicle between ultrasonography and actual size except for the size of preovulatory follicle, suggesting that information of ultrasonography is able to use for OPU or other reproductive technology of mare.

• Journal of Embryo Transfer
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• v.27 no.3
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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 $PGF_{2{\alpha}}$.

• KSBB Journal
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• v.11 no.1
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• pp.107-114
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• 1996

The enhancing effect of excreted cell factors on the production of somatic embryos from suspension cultures of Daucus carota was studied as a function of factors including molecular size, harvesting time, injection period, and concentration of the extracellular compounds. The production of late-stage embryos was increased up to 1, 500 embryos/ml compared with control cultures when high molecular size and extracellular factors, extracted from newly established embryo culture at early stationary phase, were added at the starting time. The stimulating effect on the production of somatic embryos can be attributed to the presence of high molecular size(> 10 kDa) compounds.

• Molecules and Cells
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• v.39 no.2
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• pp.136-140
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• 2016

Eph receptors and their ligands, ephrins, mediate cell-to-cell contacts in a specific brain region and their bidirectional signaling is implicated in the regulation of apoptosis during early brain development. In this report, we used the alpha(${\alpha}$)-Cre transgenic line to induce ephrin-A5 over-expression in the distal region of the neural retina. Using this double transgenic embryo, we show that the over-expression of ephrin-A5 was responsible for inducing massive apoptosis in both the nasal and temporal retinas. In addition, the number of differentiated retinal neurons with the exception of the bipolar neuron was significantly reduced, whereas the laminar organization of the mature retina remained intact. Consistent with this finding, an analysis of the mature retina revealed that the size of the whole retina-particularly the nasal and temporal regions-is markedly reduced. These results strongly suggest that the level of ephrin-A5 expression plays a role in the regulation of the size of the retinal progenitor pool in the neural retina.

• Journal of Veterinary Clinics
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• v.27 no.1
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• pp.46-49
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• 2010

In this study, we investigated the number of follicles, oocyte recovery rate and oocyte competence after in vitro maturation according to the size of follicle. And equine oocyte competence after in vitro maturation was investigated in terms of the diameter of follicle with criteria of maturation: nuclear stage after Hoechst staining. The average number of follicles per ovary with middle size (11-20 mm, 2.68) was higher than those of small (5-10 mm, 0.74) and large size follicle (> 21 mm, 1.63), therefore medium follicle (53.1%) had higher proportion than other size of follicles. The average numbers of follicle per ovary was 5.05. The rate of oocyte recovery in small (54.5%) and middle follicle (50%) was higher than that in large follicle (40.9%). After culture for 48 h in Medium 199, 50%, 45.5%, and 44.4% of oocytes from the follicles with diameters of 5-10, 11-20, > 21 mm, respectively reached the metaphase II stage. This is the first report showing number of follicle, oocyte recovery rate according to follicular size, and in vitro oocyte maturation in Jeju mare in Korea. To fulfill in vitro equine embryo production, further studies such as the seasonal effect, in vitro fertilization etc is need.

• Toxicological Research
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• v.25 no.4
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• pp.209-216
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• 2009

We have recently reported that the continuous exposure of rats to a concrete building environment under cool temperatures had adverse effects on general health parameters and embryo-fetal development. This study examined to compare the potential effects of concrete and wood building environments on pregnant dams and embryo-fetal development in rats. Groups of 10 mated females were exposed to polycarbonate (control), concrete, or wood cages from gestational days (GD) 0 to 20 under cool temperatures $(11.9\sim12.3^{\circ}C)$. All the females underwent a caesarean section on GD 20, and their fetuses were examined for any morphological abnormalities. The temperatures in the cages were similar in all groups but the relative humidity in the concrete and wood groups were higher than in the control group. The concentration of volatile organic compounds in the wood group was higher than in the control group. In the concrete group, maternal effects manifested as an increase in the incidence of clinical signs, a lower body weight, and a decrease in the thymus and ovary weights. Developmental effects included increased post-implantation loss and decreased litter size. Infrared thermal analysis showed that the skin temperature of the rats in the concrete group was lower than that in the control group. In contrast, there were no exposure-related adverse effects on the maternal and developmental parameters in the wood group. Overall, the exposure of pregnant rats to a concrete building environment under cool temperatures has adverse effects on the clinical signs, body weight, skin temperature, organ weight, and embryo-fetal development. On the other hand, exposure to a wood building environment does not have any adverse effects in rats.

• Journal of Embryo Transfer
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• v.12 no.3
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• pp.293-300
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• 1997

Supplementation of maturation medium with additional granulosa cells has beneficial effect on in vitro maturation of bovine follicular oocytes and their subsequent cleavage and development in vitro. However, maturation system using granulosa cells have some disadvantages that collection of granulosa cells is cumbersome and metabolic activity of the cells is variable according to ovarian cycle or follicular size. We hypothesized that bovine immsture oocytes matured without granulosa cell coculture can fertilize and develop normally if the medium volume per oocyte is reduced during in vitro maturation. Immature oocytes were matured for 24 hours in a TCM199 containing 10% fetal calf serum, anterior pitultary hormone (0.02 AU /ml Antrinⓡ) and estradiol with or without granulosa cells in vitro. In Group 1, 35 to 40 oocytes were matured in a well of 4-well plastic dish containing 500 $\mu$l of maturation medium and granulosa cells, and 9 to 10 oocytes were matured in a 50-$\mu$l drop of maturation medium without granulosa cells in Group 2. After maturation, oocytes were coincubated with sperm for 30 hours in a modified Tyrode's medium (IVF). Inseminated oocytes were cultured in a microdrop (30 $\mu$l) of a synthetic oviduct fluld medium (SOFM) containing BSA, Minimum Essential Medium essential and non-essential amino acids for 9 days. As a preliminary experiment, we investigated the beneficial effect of granulosa cells during maturation on subsequent cleavage and development using the same type of culturedishes (4-well dish). Granulosa cells could not increase embryo cleavage after fertilization but significantly improved (p<0.05) embryo development to expanding blastocyst (Table1 and 2). In Group 1, 68 and 80% of inseminated oocytes have cleaved at 30 hours and 2 days after IVF, respectively, which is similar (p>0.05) to the result of Group 2 (69% at 30 hours and 78% at 2 days after IVF). The oocytes in Group 2 showed 21 and 11% of developmental rates to expanding and hatching blastocysts, respectively, which was not significantly different (p>0.05) from those (20 and 10%, respectively) of oocytes in Group 1. In conclusion, it has been clarified that a microdrop culture system without granulosa cells for in vitro maturation can support bovine embryonic development to blastocyst in vitro as readily as a granulosa cell coculture system.