• Title/Summary/Keyword: Growth, Maturation

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Growth of Ovarian Primary Follicles Retrieved from Neonates of Different Ages and Derivation of Mature Oocytes Following In vitro-Culture

  • Choi, J.H.;Yoo, C.R.;Ahn, J.Y.;Park, J.H.;Lim, J.M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.5
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    • pp.629-634
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    • 2012
  • This study was conducted to improve the yield of mature oocytes from in vitro-culture of ovarian primary follicles by optimizing follicle retrieval from neonatal mice of different ages. Primary follicles of 75 to $99{\mu}m$ in diameter were collected daily from 7- to 14-day-old neonatal mice, and subsequently cultured in ${\alpha}$-MEM medium. Number of primary follicles isolated, growth of the follicle during in vitro-culture and maturation of intrafollicular oocytes were monitored. Overall, mean number of preantral follicles per animal was improved from 10.7 to 88.7 as the age of follicle donors was increased from 7 to 14-day-old. Number of primary follicles was increased gradually up to 11-day-old (35.7 follicle per an animal), then reduced to 29 in 14-day-old (p = 0.0013). More follicles retrieved from 10-day-old or 11-day-old females maintained their morphological normality at the end of primary culture than the follicles retrieved from 9-day-old. Of those cultured, primary follicles retrieved from 11-day-old mice yielded largest larger number of early secondary follicles than the follicles retrieved from in the other ages (39 vs. 13 to 29%). More than 3.3-times increase (0.86 to 2.86; p<0.05) in an average number of mature oocytes per animal was observed in the group of 11-day-old, compared with 9-day-old. However, no difference was found in the percentage of primary follicles developing into the pseudoantral stage (21 to 30%; p = 0.5222) and in the percentage of oocytes mucified (32 to 39%; p = 0.5792). In conclusion, a positive correlation between retrieval time and follicle growth was detected, which influences the efficiency to derive mature oocytes by follicle culture.

IMMUNOHISTOCHEMICAL PROFILE OF BASIC FIBROBLAST GROWTH FACTOR(bFGF) IN GROWING RAT T-M JOINT (성장 중인 흰쥐 악관절의 basic fibroblast growth factor(bFGF) 분포에 관한 면역조직화학적 연구)

  • Kim, Jong-Ha;Lee, Sang-Chull
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.27 no.1
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    • pp.1-8
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    • 2001
  • This study was designed to localize the distribution of basic fibroblast growth factor(bFGF) in the developing rat condylar region and to elucidate the associated function of bFGF in the condyle development. The condyles of temporomandibular joint of Sprague-Dawley rats (27g of weight) were used. The tissues were examined with electron microscope and immunohistochemical method. The results were as follows: 1. The developing condylar region are divided in to 5 zones apparently: proliferative, maturation, hypertrophic, calcifying, and ossification zones. 2. The cells in the proliferative zone are condensed and have under-developed cell organells in the cytoplasm. This zone shows a strong immunoreactivity of bFGF. 3. The cells in the maturation zone are typical chondroblasts showing well-developed cell organells and round nucleus. The cartilaginous matrix does not show the immunoreactivity of bFGF, while the chondroblasts show the immunoreactivity. 4. The cells in the hypertrophic zone show hypertrophic change having the degenerated cell organelles and small nucleus. There are no immunoreactivity of bFGF in this zone except the nucleus and endoplasmic region showing mild immunoreactivity. 5, The cells in the calcifying zone show hypertrophic change and cell organelles are disappeared. The cells are surrounded by the calcified cartilaginous matrix. There are no immunoreactivity of bFGF in this zone except the endoplasmic region showing mild immunoreactivity. 6. In the zone of bone formation, chondroblasts are disappeared. Newly differentiated osteoblasts secreting osteoid around the calcified cartilaginous matrix. The bone marrow shows the immunoreactivity of bFGF, while the bone matrix does not show the immunoreactivity of bFGF.

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Induction of Sex Maturation and Growth in Comb Pen Shells, Atrina pectinata per Microalgae Classes (미세조류 종류에 따른 키조개, Atrina pectinata의 성장 및 성숙 유도)

  • Moon, Tae-Seok;Jo, Pil-Gue;Kim, Byoung-Hak;Park, Ki-Yeol;Ku, Hag-Dong;Shin, Yun-Kyung;Lym, Young-Sub
    • The Korean Journal of Malacology
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    • v.25 no.2
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    • pp.105-112
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    • 2009
  • We investigated the degree of obesity, histological development stages of gonads and sexual maturation induction rates of comb pen shell, Atrina pectinata, per the type of micro-algae supplied. Terms of maturation by singular or mixed supply of microalgae, it was found that maturation of the female was the quickest at 60.0% by the Tetraselmis tetrathele (Tet). experiment group followed by 57.1% by the Chlorella ellipsoidea (Chl). experiment group and 16.7% by the Phaeodactylum tricornutum (Pha). experiment group. However, there were no significant differences between Tet. experiment group and Chl. experiment group. As for the male, maturation was the quickest at 60.0% by the Tet. experiment group followed by 16.7% by the Chl. experiment group and 14.3% by the Pha. experiment group. In light of these results, Tet. is concluded to be a very useful feed organism in breeding the mother comb pen shells. Upon completion of the experiment, the sexual maturation induction rate for the female was found to be the highest at 82.0% in the Tet. experiment group followed by 72.0% by the Chl. experiment group, 64.0% by the Pha. experiment group and 58.0% by the mixed micro-algae experiment group. During the period of experiment, the survival rate was the highest at 94.4% by the mixed micro-algae experiment group followed by 90.0% by the Pha. experiment group, 83.1% by the Tet. experiment group and 78.8% by the Chl. experiment group.

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Estimation of Growth Curve Parameters for Body Weight and Length in Miniature Pigs

  • Kang, Hyun Sung;Nam, Ki Chang;Cabling, Meriam M.;Lee, Myeong Seop;Choi, Te Jung;Yoon, Jong Taek;Seo, Kang Seok
    • Journal of Animal Science and Technology
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    • v.54 no.6
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    • pp.395-400
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    • 2012
  • This study was conducted to estimate the growth curve parameters for the body weight (BW) and body length (BL) of miniature pigs in Korea. Growth curve parameters were estimated through a nonlinear regression model using Gompertz, Logistic, and von Bertalanffy methods. A total of 25 piglets were measured monthly from birth up to 15 months of age to estimate both body weight and length. Results showed that the estimated average values for the body weight (body length) were 31.83 kg (58.77 cm) for the mature weight (A), 3.06 (1.74) for the growth ratio (${\beta}$), and 0.28 (0.52) for the maturing rate (${\kappa}$). Average inflection points showing maximum growth rate estimated each month for body weight were 3.97 kg and 11.70 cm, while for the body length were 1.06 kg and 21.61 cm. Moreover, the estimated maturation rates of the body weight and length for the group of Sire 1 were 0.22 and 0.40 respectively, whereas for the group of Sire 2 these values were 0.34 and 0.39. On the other hand, for the groups of Dam 1, Dam 2, and Dam 3, maturation rates for their body weights were 0.26, 0.28 and 0.33 respectively, while for their body lengths these values were 0.43, 0.37, and 0.38, respectively. The study also indicated a negative relationship between the values of mature weight and maturity rate for the body weight will result to a higher inflection point which is in contrast for the body length where results show that a positive relationship between the values of mature length and the maturity rate will result to a higher inflection point. Furthermore, the growth performance of miniature pig varies across stages but using these estimated growth curve parameters could improve the genetic traits of miniature pig.

Change in chemical composition of maturing barley kernels (보리곡립의 성숙중 이화학적 특성변화)

  • Seog, Ho-Moon;Kim, Jong-Sang;Hong, Hee-Do;Kim, Sung-Soo;Kim, Kyung-Tack
    • Applied Biological Chemistry
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    • v.36 no.6
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    • pp.449-455
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    • 1993
  • Some chemical properties were investigated for barley kernels at different growth stages. Crude fat, crude protein, starch, alcohol-insoluble solids(AIS), and ${\beta}-glucan$ increased until $31{\sim}36\;days$ from ear emergence and thereafter remained relatively constant, whereas ash and crude fiber contents were gradually decreased through the whole period of maturation. As barley kernels became mature, their water contents kept decreasing, and the content of milky stage barley kernel at $31{\sim}36th\;days$ from ear emergence ranged from $40{\sim}50%$. Free sugars including glucose, fructose, sucrose, raffinose, maltose, and kestose were identified in the growing kernels. Glucose, sucrose and kestose decreased with maturation while raffinose slightly increased. Barley kernels on the 43rd day from ear emergence contained 0.62% sucrose, 0.46% raffinose, 0.33% kestose, 0.19% glucose, 0.17% fructose, 0.04% maltose. Analysis of minerals for barley kernels at different growth stages showed that the growing kernels contained K, P, Mg, Ca, Na, Fe, Zn, Mn, Cu in decreasing order. Vitamin $B_1$ content tended to increase with kernel growth, showing maximum value of $350\;{\mu}g\;per\;100\;g$ dry weight on 36th day from ear emergence.

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Effect of Ethephon and Dichlorprop on the Growth and Maturation of Fruit in 'Niitaka' Pears (Ethephon 및 Dichlorprop 처리가 배 '신고'의 과실 생장과 성숙에 미치는 영향)

  • Lee, Jae Chang;Kwon, Oh Won;Hwang, Yong Soo
    • Korean Journal of Agricultural Science
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    • v.21 no.2
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    • pp.81-91
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    • 1994
  • This study was conducted to find a potential effect of ethephon or dichlorprop spray on the fruit growth and maturation in 'Niitaka' pears. Chemicals were applied between June 10 to 24 (7-9 weeks after full bloom). Ethephon in the range from 25 ppm to 100 ppm, and dichlorprop from 20 ppm to 40 ppm reduced the fruit size regardless application time and concentration. The fruit growth was more inhibited at the higher concentrations in both chemicals. Flesh firmness was more rapidly decreased in the fruit treated by both chemicals. Firmness at harvest was lowest in the treatment of 100 ppm ethephon. The climacteric increase of ethylene synthesis occurred earlier in the fruit treated by ethephon or dichlorprop regardless application concentrations and the maximum peak of ethylene production was rugher. Total phenolics significantly reduced in fruits treated with ethephon. However, the contents of soluble solids and acid were not affected by ethephon or dichlorprop treatment. Ground color was changed rapidlys by the treatments resulting in the early maturation of fruit. Optimum maturity of fruit was shortened 2-3 days by ethephon and 3-4 days by dichlorporp when maturity was determined on the basis of ground color development and firmness.

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In Vitro Maturation, Fertilization and Development of Mouse Oocytes Derived from In Vitro Grown Preantyal Follicles (체외성장된 Preantral Follicle에서 유래된 생쥐난자의 체외성숙, 수정 및 발달)

  • Kim, D.H.;Lee, H.J.;K.S. Chung;Lee, H.T.
    • Korean Journal of Animal Reproduction
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    • v.26 no.2
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    • pp.133-142
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    • 2002
  • The objective of this study was to examine maturation, fertilization and developmental rate of the in vitro-grown mouse oocytes, and to compare these results with those of oocytes grown and matured in vivo. The preantral follicles isolated from 12-day-old mice were cultured on Transwell-COL membrane inserts. After in vitro growth and maturation, 72.5 % of oocytes grown in vitro produced polar body which can be comparable to in vivo growth (70.5 %). However, the mean oocyte diameter of the in vitro group (69.6$\pm$2.1$\mu$m) was smaller than that of the in vivo group (73.3$\pm$3.0$\mu$m). The fertilization rate was significantly lower (p<0.05) in the in vitro group (76.5%) than in the in vivo group (90.2%), however, there was no difference in the percentage of monospermic and polyspermic oocytes between two groups. The capacities of in vitro grown ova to cleave and develop to blastocyst were (57.8 and 14.4%, respectively) significantly lower (p<0.001) than those of the in vivo counterpart (84.4 and 56.6%, respectively). Moreover, the mean number of cells per blastocyst was significantly lower (p<0.05) in the in vitro group (39.0$\pm$10.8) than in the in vivo group (60.5$\pm$12.5). Live young were produced from transferred 2-cell embryos derived from in vitro-grown and matured oocytes. In conclusion, the results show that in vitro-grown oocytes did not achieve the developmental capacity of in vitro-grown oocytes.

Effect of Epidermal Growth Factor (EGF) on Meiotic Maturation and Pronuclear Formation of Porcine Oocytes Produced In Vitro

  • Song S. H.;Kim J. G.;Song H. J.;Kumar B. Mohana;Cho S. R.;Choe C. Y.;Choi S. H.;Rho G. J.;Choe S. Y.
    • Reproductive and Developmental Biology
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    • v.29 no.2
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    • pp.127-131
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    • 2005
  • The objective of this study was to examine the effect of EGF on meiotic maturation and pronuclear (PN) formation of porcine oocytes. Prepubertal gilt cumulus-oocyte-complexes (COCs) aspirated from $2\~6mm$ follicles of abbatoir ovaries were matured in TCM199 containing 0.1mg/ml cysteine, $0.5{\mu}/ml$ FSH and LH, and EGF (0, 5, 10, 20, 40 ng/ml) for 22 hr at $39^{\circ}C$ in a humidified atmosphere of $5\%$ $CO_2$ in air. They were then cultured for an additional 22hr without hormones. In Experiment 1, to examine the nuclear maturation at 44hr of culture, the expanded cumulus cells were removed by vortexing for 1 min in 3 mg/ml hyaluronidase. The oocytes were fixed in acetic acid: methanol (1:3, v/v) at least for 48 hr and stained with $1\%$ orcein solution for 5 min. Nuclear status was classified as germinal vesicle (GV), germinal vesicle breakdown (GVBD), prophase-metaphase I (PI-MI), and PII-MII under microscope. In Experiment 2, to investigate PN formation, oocytes were fertilized with Percoll-treated freshly ejaculated sperm $(1\times10^5\; cells/ml)$ in mTBM with $0.3\%$ BSA and 2mM caffeine for 5hr, and cultured in NCSU-23 medium with $0.4\%$ BSA. At 6hr of culture, the embryos were fixed in $3.7\%$ formaldehyde for 48hr and stained with 10ug/ml propidium iodide for 30 min. PN status was classified as no or one PN (unfertilized), 2 PN (normal fertilized) and $\geq3$ PN (polyspermy). Differences between groups were analyzed using one-way ANOVA after arc-sine transformation of the proportional data. The rate of oocytes that had reached to PII-MII were significantly (P<0.05) higher in all groups added EGF than that of non-treated group $(67\%)$, but it did not differ among the all added groups $(86\%,\;85\%,\;79\%\;and\;81\%$, in 5, 10, 20 and 40 ng/ml EGF, respectively). No differences on the incidence of 2PN were observed in all treated groups $(25\%,\;30\%,\;33\%,\;29\%\;and\;29\%$, in 0, 5, 10, 20 and 40 ng/ml EGF, respectively), however, in non-treated group, polyspermy tended to be increased ($66\%\;vs\;. 58\%,\;54\%,\;52\%\;and\;55\%$, 0 vs. 5, 10, 20, 40 ng/ml EGF, respectively). These results suggest that EGF can be effectively used as an additive for enhancing oocyte maturation and reducing the incidence of polyspermy in pig.

Glutathione (GSH) Concentration and Developmental Competence of Korean Native Cow Oocytes Selected by Brilliant Cresyl Blue (BCB)

  • Lee, Hyo-Sang;Yu, Dae-Jung;Kwon, Tae-Hyeon;Cho, Su-Jin;Bang, Jae-Il;Park, Sang-Guk;Cho, Sung-Kyun;Kong, Il-Keun
    • Journal of Embryo Transfer
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    • v.24 no.1
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    • pp.39-45
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    • 2009
  • This study was carried out to evaluate the nuclear, cytoplasmic maturation and developmental potential of bovine oocytes selected by brilliant cresyl blue (BCB) as indirect measurement of oocytes growth phase. Cumulus-oocyte complexes (COCs) were collected from 2 to 8 mm follicles from slaughterhouse Hanwoo ovaries. The COCs were divided into stained cytoplasm to blue (BCB+) and unstained (BCB-) according to their ooplasm BCB coloration stained by $26{\mu}m$ of BCB after 90 min. Selected COCs were cultured in a TCM 199 for 18 to 26 h. Nuclear maturation and total cell number was evaluated after in vitro maturation (IVM) or in vitro culture (IVC) using $10{\mu}g/ml$ Hoechst 33342, and cytoplasmic maturation was evaluated by intracellular glutathione (GSH) assay before (0 h) and after (24 h) IVM. The oocyte diameters were not differed significantly between BCB+ ($157.4{\pm}5.8{\mu}m$) and BCB+ ($149.0{\pm}31.0{\mu}m$) groups (p>0.05). However, the proportion of metaphase II oocytes in BCB+ group was significantly higher than BCB- group after IVM (p<0.05). GSH content of BCB+ group oocytes was significantly higher than that of BCB- group just after collection ($7.3{\pm}0.6$ vs. $4.8{\pm}0.6\;pmol/oocyte$, p<0.05), but not varied after IVM($13.1{\pm}0.9$ and $12.6{\pm}2.5\;pmol/oocytes$ for BCB+ and BCB- respectively; p>0.05). The proportion of blastocyst formation and total cell number in BCB+ group (23.5% and $105.5{\pm}28.6$) was significantly higher than that in BCB- (9.8% and $72.4{\pm}26.1$; p<0.05). The results indicate that BCB+ group oocytes may provide a cellular and functional basis for the greater developmental competence in Korean Native Cow (KNC) oocytes.