• Title/Summary/Keyword: Medium development

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Effect of MEM Vitamins Supplementation of In vitro Maturation Medium and In vitro Culture Medium on the Development of Porcine Embryos

  • Kim, J.Y.;Lee, E.J.;Park, J.M.;Park, H.D.
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.11
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    • pp.1541-1546
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    • 2011
  • This study was carried out to examine the influence of minimum essential medium (MEM) vitamins supplementation to in vitro maturation medium and in vitro culture medium on the development of porcine embryos. Porcine embryo development was investigated following cultivation in both in vitro maturation and culture medium with the supplementation of MEM vitamins (0, 0.1, 0.2 and 0.4%) using immature oocytes collected from the ovary of prepubertal gilts. Embryo development was observed and the total cell number in each blastocyst generated under the culture conditions was quantified following supplementation of the medium. The embryonic development rate of the blastocyst and hatched blastocyst was higher, but not significantly so, when 0.4% MEM vitamins were supplemented to the in vitro maturation medium of the porcine oocyte. Interestingly, the total number of cells in the blastocyst was significantly higher in the in vitro maturation MEM vitamins supplemented group compared to either the untreated group or the group which had MEM vitamins supplemented to both in vitro maturation and in vitro culture medium simultaneously (p<0.05). Therefore, the supplementation of 0.4% MEM vitamins to the in vitro mature medium has a beneficial effect on the embryonic development of in vitro produced blastocysts derived from the immature porcine oocytes.

Study on Development of In Vitro Culture Medium for Rabbit Embryos (토끼 수정란 체외 배양액의 개발에 관한 연구)

  • 임경순;진동일;김대경;김성우;정소용;최화식
    • Korean Journal of Animal Reproduction
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    • v.22 no.1
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    • pp.35-42
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    • 1998
  • This experiment was carried out to improve in vitro development of rabbit one-cell embryos to the blastocyst stage. One-cell rabbit embryos were collected at 19\ulcorner20hr after superovulation induction and incubated at 39\ulcorner in 5% CO2 for 72hr. In order to find optimum conditions in medium that affects the rabbit embryo's development in vitro, RDH medium which mixed with RPMI1640, DMEM and Ham's F10 was compared with the previously reported mediums (Ham's F10 and RD) for embryo development and cell numbers. Three additives (BSA, taurine and glucose) were tested for the development of rabbit one-cell embryos in vitro. When the embryos were cultured in RDH medium, their development was markedly promoted as compared with Ham's F-10 or RD alone. Glucose exhibited no significant effects on embryo development and cell numbers. BSA a, pp.ared to promote transition from morula to blastocyst stage and taurine increased cell numbers of cultured embryos markedly regardless of medium. BSA and taurine together in RDH medium showed the additive effects on embryos development and cell number.

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Effect of Glucose and Sodium Phosphate on In Vitro Development of Porcine Embryos

  • Lee, S.H.;Lim, S.M.;Lee, S.Y.;Cheong, H.T.;Yang, B.K.;Park, C.K.
    • Reproductive and Developmental Biology
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    • v.28 no.2
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    • pp.101-105
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    • 2004
  • This study was carried out to evaluate the effects of glucose and sodium phosphate on in vitro development of porcine oocytes matured and fertilized in vitro. When the culture medium was supplemented with various concentrations of glucose, the higher proportions (23 and 26%) of oocytes developed to morular or blastocyst stages were at the concentrations of 2.78 and 5.56 mM than 0 (9%; P<0.05) and 11.12 mM (18%). In experiment to evaluate effect of sodium phosphate during in vitro development of porcine oocytes, a significantly (P<0.05) higher proportions of embryos developed to morular or blastocyst stages was obtained with sodium phosphateof 0.28 (25%) and 0.53 (27%) mM than 0 (15%), 1.05 (19%) and 2.10 (10%) mM. On the other hand, when oocytes were cultured in medium with (0.53 mM) sodium phosphate, the proportions of developed embryos were significantly (P<0.05) higher in medium without (29%) that than with (14%) 5.56 mM glucose. However, a higher proportion of embryos developed to morular or blastocyst stages were obtained in medium with (23%) that than without (8%) glucose (P<0.05). The minimum essential medium (MEM) added to the culture medium were higher regardless of presence of sodium phosphate and glucose on the development of embryos. Although sodium phosphate and glucose could support morular and blastocyst development to a limited extend (10∼24%), significantly higher proportion (36%) at morular or blastocyst stages was obtained by MEM adding in the medium with sodium phosphate and glucose. These results suggest that the early development of in vitro fertilized porcine oocytes can be maintained efficiently by glucose and sodium phosphate when they were cultured in medium with MEM.

In Vitro Fertilization and Development of Mouse Eggs (생쥐난자의 시험관내 수정과 발달)

  • 김승재;정길생
    • Korean Journal of Animal Reproduction
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    • v.8 no.2
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    • pp.110-115
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    • 1984
  • These experiments were carried out to obtain the information about the optimal pH osmolality affecting in vitro fertilization of the mouse eggs, to elucidate the 2-cell block to development in vitro and to find out the method of controlling the subsequent embryo development in vitro. pH and osomlality was adjusted by adding NaCl or NaHCO3 to the basic salt solution. In vitro fertilization were carried out by inroducting the cumulus masses to the suspension of epididymal spermatozoa at each pH, osmolality, and 10${\mu}$M-EDTA medium. The results obtained in these experiments were summarized as follows: 1. The fertilization rates in vitro at each medium of 235, 252, 269, 286, 306, 323, 345, 368, 393 mosmol were 15.6, 38.2, 65.7, 75.6, 80.9, 74.3, 58.1, 35.1, 24.3, 11.1%, respectively. 2. The fertilization rates in vitro at each medium of pH 6.1, 6.4, 6.7, 7.0, 7.3, 7.6, 7.9, 8.1 were 11.8, 17.9, 32.4, 61.9, 79.5, 76.7, 53.5, 13.6%, respectively. 3. In case of ICR female x ICR male embryos, the development rate of 2-cell embryos to 4-8 cell embryos was 16.2% at normal medium, but the rate was increased to 49.3% in medium containging 10 ${\mu}$M-DETA; In case of C3H female x ICR male embryos, the development rate was 41.0% at normal medium, but the rate was increased to 71.7% at 10 ${\mu}$M-EDTA-medium.

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Effect of Oviductal Fluid and Oviductal Conditioned Medium on Polyspermy and In Vitro Development of Porcine Oocytes (돼지 난관액과 Oviductal Conditioned Medium 이 다정자침입과 체외배발달에 미치는 영향)

  • 문승주;김재홍;나진수
    • Korean Journal of Animal Reproduction
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    • v.22 no.4
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    • pp.411-417
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    • 1998
  • The objective of this study was to dertermine the effects of oviductal fluid and oviductal conditioned medium on polyspermy and in vitro development of porcine oocytes. The addition of oviductal fluid and oviductal conditioned medium in the prefertilization and fertilization medium significantly decreased polyspermy rates and the mean number of spermatozoa in penetrated eggs(P<0.05). The acrosome reaction rate significantly increased when spematozoa were exposed for 1.5, 3, 4.5h in oviductal fluid and oviductal conditioned medium(P<0.05). When oocytes cultured for 192h, the percentage of oocytes that developed to the morula and blastocyst stage was higher in culture medium with oviductal fluid and oviductal conditioned medium than without oviductal fluid and oviductal conditioned medium(P<0.05). These results indicated that the oviductal secreations will effectively reduce both the polyspermy rates and the mean number of spermatwa in penetrated eggs. And the presence of culture with oviductal fluid and oviductal conditioned medium promotes in vitro development of porcine oocytes.

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Recycling Post-harvest Medium from Bottle Cultivation for Oyster mushroom(Pleurotus ostreatus) (버섯 병재배 수확후배지의 느타리버섯 배지에 알맞은 재활용 수준)

  • Cheong, Jong-Chun;Lee, Chan-Jung;Shin, Pyung-Gyun;Suh, Jang-Sun
    • Journal of Mushroom
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    • v.10 no.4
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    • pp.167-173
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    • 2012
  • This study was carried out to test the recycling post-harvest medium from other mushroom bottle cultivation as a secondary medium of the oyster mushroom. In the post-harvest medium from winter mushroom and king oyster mushroom cultivation, oyster mushroom varieties in Chunchu-2ho and Manchuri fruit bodies yields compared with control group tend to be low. After recycling the post-harvest medium, it was replaced by basal medium up to 50%, of which the fruit bodies with stable yield increase from 10% to 30% were increased.

Development of Cheap Substrate for Fruiting of Pleurotus ostreatus using Paper Sludge (제지 부산물을 이용한 느타리버섯(Pleurotus ostreatus) 자실체 형성용 염가배지개발)

  • Jo, Woo-Sik;Yun, Yeong-Seok;Park, Sun-Do;Choi, Boo-Sull
    • The Korean Journal of Mycology
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    • v.23 no.3 s.74
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    • pp.197-201
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    • 1995
  • For 2 years $(1993{\sim}1994)$, study on development of cheap medium for Pleurotus ostreatus revealed that paper sludge contain more CaO and similar T-C, T-N, $P_2O_5$, MgO but less $K_2O$ than any other medium material in chemical property. Mixed treatment (rice straw + paper sludge 10, 30, 50%, cotton waste + paper sludge 10, 30, 50%, cotton waste + rice hull 20 + paper sludge 10, 30, 50, 70%) is similar or fast a little in mycelial growth and is similar or fast $1{\sim}2$ day in period of primordia formation than cotton waste medium, and in the yield to each medium type also increased but excepted in rice hull 20% + paper sludge 70%, especially mixed medium at 7:3 ratio of cotton waste and paper sludge is best treatment because it is increased to 21%. In economical analysis, mixed medium at 7:3 ratio of cotton waste and paper sludge is increased to 50% compared to cotton waste medium in relative income.

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Plant Regeneration from Shoot Tip-Derived Embryogenic Callus of Dianthus superbus

  • Lee, Eun-Ae;Kim, Joon-Chul;Kim, Won-Bae;Kim, Byeong-Hyeon;Kim, Jeong-Kan
    • Journal of Plant Biology
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    • v.37 no.3
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    • pp.381-385
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    • 1994
  • The highest degree of callus formation was obtained from the shoot tips of Dianthus superbus when cultured on the MS medium supplemented with 2.0 mg/L NAA and 0.5 mg/L BAP. Embryogenic calluses were obtained from the seperated friable calluses on MS medium containing 2.0 mg/L 2,4-D after 7-8 wk of culture. For plant regeneration, embryogenic calluses were selected and cultured on te proliferation medium. After 3 wk, somatic embryos appeared on MSK medium (0.5 mg/L NAA, 2.0 mg/L kinetin) and N6 medium (2.0 mg/L kinetin, 0.1 mg/LNAA, 0.1 mg/L 2,4-D and 2.0 g/L casein hydrolysate). When these somatic embryos were kept under continuous illumination, shoots were successfully regenerated on the both media. The shoots were rooted on MS medium supplemented with 2.0 mg/L NAA.

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Conditioned medium of E17 rat brain cells induced differentiation of primary colony of mice blastocyst into neuron-like cells

  • Budiariati, Vista;Rinendyaputri, Ratih;Noviantari, Ariyani;Haq, Noer Muhammad Dliyaul;Budiono, Dwi;Pristihadi, Diah Nugrahani;Juliandi, Berry;Fahrudin, Mokhamad;Boediono, Arief
    • Journal of Veterinary Science
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    • v.22 no.6
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    • pp.86.1-86.13
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    • 2021
  • Background: Conditioned medium is the medium obtained from certain cultured cells and contained secretome from the cells. The secretome, which can be in the form of growth factors, cytokines, exosomes, or other proteins secreted by the cells, can induce the differentiation of cells that still have pluripotent or multipotent properties. Objectives: This study examined the effects of conditioned medium derived from E17 rat brain cells on cells with pluripotent properties. Methods: The conditioned medium used in this study originated from E17 rat brain cells. The CM was used to induce the differentiation of primary colonies of mice blastocysts. Primary colonies were stained with alkaline phosphatase to analyze the pluripotency. The morphological changes in the colonies were examined, and the colonies were stained with GFAP and Neu-N markers on days two and seven after adding the conditioned medium. Results: The conditioned medium could differentiate the primary colony, beginning with the formation of embryoid-body-like structure; round GFAP positive cells were identified. Finally, neuron-like cells testing positive for Neu-N were observed on the seventh day after adding the conditioned medium. Conclusions: Conditioned medium from different species, in this case, E17 rat brain cells, induced and promoted the differentiation of the primary colony from mice blastocysts into neuron-like cells. The addition of CM mediated neurite growth in the differentiation process.

Effects of astaxanthin supplementation in fertilization medium and/or culture medium on the fertilization and development of mouse oocytes

  • Tana, Chonthicha;Somsak, Pareeya;Piromlertamorn, Waraporn;Sanmee, Usanee
    • Clinical and Experimental Reproductive Medicine
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    • v.49 no.1
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    • pp.26-32
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    • 2022
  • Objective: We investigated the effect of supplementing fertilization medium and/or culture medium with astaxanthin (AST) on the two phases of in vitro fertilization: gamete fertilization and embryo development. Methods: Mouse cumulus-oocyte complexes were divided into four groups with 5 µM AST added to the fertilization medium (group 3, n=300), culture medium (group 2, n=300), or both media (group 4, n=290). No AST was added to the control group (group 1, n=300). Results: The fertilization rate was significantly higher (p<0.001) in the groups using AST supplemented fertilization medium (group 3, 79.0%; group 4, 81.4%) than those without AST (group 1, 56.3%; group 2, 52.3%). The blastocyst rate calculated from the two-cell stage was significantly lower (p<0.001) in the groups using AST-supplemented embryo culture medium (group 2, 58.0%; group 4, 62.3%) than in those without AST (group 1, 82.8%; group 3, 79.8%). The blastocyst rate calculated from the number of inseminated oocytes was highest in group 3 (189/300, 63.0%) and lowest in group 2 (91/300, 30.3%) with statistical significance compared to other groups (p<0.001). There were significantly higher numbers of cells in the inner cell mass and trophectoderm, as well as significantly higher total blastocyst cell counts, in group 3 than in the control group. Conclusion: An increased blastocyst formation rate and high-quality blastocysts were found only in the fertilization medium that had been supplemented with AST. In contrast, AST supplementation of the embryo culture medium was found to impair embryo development.