scholarly journals Penambahan Glutathione pada Medium Fertilisasi Efektif Mendukung Pembentukan Pronukleus dan Perkembangan Awal Embrio Sapi (SUPPLEMENTATION OF GLUTATHIONE IN FERTILIZATION MEDIUM EFFECTIVELY SUPPORT NORMAL PRONUCLEUS FORMATION AND EARLY BOVINE EMBRYONIC DE

2017 ◽  
Vol 18 (3) ◽  
pp. 327
Author(s):  
Aras Prasetiyo Nugroho ◽  
Iman Supriatna ◽  
Mohamad Agus Setiadi
Keyword(s):  
Tissue Culture ◽  
Embryonic Development ◽  
Culture Medium ◽  
Fertilization Rate ◽  
Tissue Culture Medium ◽  
Randomized Design ◽  
Oviduct Fluid ◽  
And Control ◽  
Bovine Oocytes

The objective of this study was to determine fertilization rate effectiveness and early embryonic development competency with glutathione (GSH) supplementation in fertilization medium and culture This study consisted of two experiments comprising each of the four treatment and six repetitions with completely randomized design (CRD) using 651 oocytes. In the first experiment, a total of 317 bovine oocytes were matured in tissue culture medium (TCM) 199 at incubator 5% CO2 with temperature 39 ºC for24 h, then fertilized with sperm separated by swim up technique. Oocyte and sperms were incubated in fertilization medium supplemented with 0.25 mM, 0.50 mM, 1.00 mM GSH. In the second experiment, bovine oocytes were matured in maturation medium and fertilized with same procedure as mentioned before, then cultured in modified synthetic oviduct fluid (mSOF) with the following treatment: supplementation GSH only in fertilization medium (T1), supplementation GSH only in culture medium (T2), and supplementation GSH in both fertilization and culture medium (T3), while control not supplementation GSH (T0). Result of the first experiment showed that supplementation 1.00 mM GSH in fertilization medium can increase higher normal pronucleus (2PN) formation (86,9%) compared to other treatments, 0.50 mM (80.3%), 0.25 mM (73.8%), and control (58.9%) (P<0.05). In the second experiment showed that early bovine embryonic development on 2nd day cultured which reached 5-8 cell on treatment T1 (56.0%) and T3 (53.6%) were higher (P<0.05) compared to treatment T2 (26.2%) and T0 (control) (31.3%). Result of the other were showed that early bovine embryonic development on 4th day cultured which reached 9-16 cell on treatment T1 (26.2%) and T3 (27.4%) were higher (P<0.05) compared to that T2 (11.9%) and T0 (control) (10.8%). In conclusion, 1.00 mM GSH supplementation in medium was more effective in supporting normal pronucleus formation and early fertilization bovine embryonic development compared to in culture medium. ABSTRAK Penelitian ini bertujuan untuk mengetahui tingkat fertilisasi dan kompetensi perkembangan awal embrio sapi dengan penambahan glutathione (GSH) pada medium fertilisasi in vitro (IVF) dan kultur in vitro (IVC). Penelitian ini terdiri atas dua penelitian yang terdiri dari masing-masing empat perlakukan dan enam kali ulangan dengan rancangan acak lengkap (RAL) menggunakan 651 oosit. Penelitian I, sebanyak 317 oosit sapi dalam tissue culture medium (TCM) 199 dimatangkan pada inkubator 5% CO2 dan suhu 39°C selama 24 jam, kemudian difertilisasi dengan spermatozoa yang telah diseleksi menggunakan teknik swim up. Oosit dan spermatozoa diinkubasi pada medium fertilisasi dengan penambahan 0,25 mM, 0,50 mM, dan 1,00 mM GSH. Penelitian II, sebanyak 334 oosit sapi dimatangkan pada medium pematangan dan difertilisasi, kemudian dikultur pada medium modified synthetic oviduct fluid (mSOF), dengan perlakuan: penambahan GSH hanya pada medium fertilisasi (T1), penambahan GSH hanya pada medium kultur (T2), dan kombinasi penambahan GSH pada medium fertilisasi dan kultur (T3). Hasil penelitian I, menunjukkan bahwa penambahan 1,00 mM GSH pada medium fertilisasi dapat meningkatkan pembentukan pronukleus normal (2PN) yang lebih tinggi (86,9%) dibandingkan dengan perlakuan yang lain yaitu 0,50 mM (80,3%), 0,25 mM (73,8%), dan kontrol (58,9%) (P<0,05). Penelitian II menujukkan bahwa perkembangan awal embrio sapi pada hari ke-2 kultur yang mencapai pembelahan 5-8 sel pada perlakukan T1 (56,0%) dan T3 (53,6%) lebih tinggi (P<0,05) dibandingkan dengan perlakuan T2 (26,2%) dan T0 (kontrol) (31,3%). Hasil penelitian lain menunjukkan bahwa perkembangan awal embrio sapi pada hari ke-4 kultur yang mencapai pembelahan 9-16 sel pada perlakuan T1 (26,2%) dan T3 (27,4%) lebih tinggi dibandingkan dengan perlakukan T2 (11,9%) dan T0 (kontrol) (10,8%) (P<0,05). Dapat disimpulkan bahwa penambahan 1,00 mM GSH pada medium fertilisasi lebih efektif dalam mendukung pembentukan pronukleus normal dan perkembangan awal embrio sapi dibandingkan pada medium kultur.

EVIDENCE FOR THE INDUCTION OF URIDINE AND DEOXY-URIDINE PHOSPHORYLASES OF REGENERATING RAT LIVER IN VITRO

1965 ◽  
Vol 43 (1) ◽  
pp. 41-48 ◽  
Author(s):  
Esther W. Yamada
Keyword(s):  
Amino Acids ◽  
Tissue Culture ◽  
Rat Liver ◽  
Culture Medium ◽  
Specific Activity ◽  
Tissue Culture Medium ◽  
Uridine Phosphorylase ◽  
Regenerating Rat Liver ◽  
Specific Activities

Increases in the specific activities of undine and deoxyuridine phosphorylases of slices of regenerating rat liver were found 4 hours after incubation in tissue-culture medium containing uridine or 6-azauridine. These increases were not found when the tissue-culture medium contained either 8-azaguanine or puromycin, or when it lacked amino acids. Although both uridine and 6-azauridine were more effective in increasing the specific activity of uridine phosphorylase than that of deoxyuridine phosphorylase, azauridine was more effective than uridine in increasing the specific activities of both enzymes.In time studies, in which slices of regenerating rat liver were incubated in tissue-culture medium containing optimal concentrations of uridine, the specific activities of the two enzymes reached maximum levels at 3–4 hours. Puromycin prevented these increases.

scholarly journals A Method for Chronological Intravital Imaging of Bovine Oocytes during In Vitro Maturation

2008 ◽  
Vol 14 (6) ◽  
pp. 549-560 ◽  
Author(s):  
Morten R. Petersen ◽  
Michael Hansen ◽  
Birthe Avery ◽  
Ingrid B. Bøgh
Keyword(s):  
Embryonic Development ◽  
Oocyte Maturation ◽  
Laser Scanning ◽  
Subsequent Development ◽  
Cumulus Cells ◽  
Fertilization Rate ◽  
Laser Scanning Microscopy ◽  
Intravital Imaging ◽  
Bovine Oocytes

AbstractOocyte maturation is known to affect the chances for successful fertilization, embryonic development, establishment of pregnancy and delivery of a live, healthy, and viable offspring. Two-photon laser scanning microscopy (TPLSM) has previously been used to evaluate early embryonic development without a detectable impairment of subsequent development, but has never been applied to assess mammalian oocytes throughout in vitro maturation (IVM). Visualization of structures within live oocytes during IVM, followed by fertilization and embryo culture, may improve the understanding of oocyte maturation. To visualize structures within bovine oocytes using TPLSM, it is necessary to remove the cumulus cells that normally surround the oocyte during maturation. Repeated visualization of structures within the same oocyte is possible, if movement of the oocyte can be avoided. In this article, we describe the development of a method for repeated intravital imaging of denuded bovine oocytes using an upright TPLSM equipped with a specially constructed incubator. Oocytes were stained with Hoechst 33258, and the nuclear structures were evaluated. Oocyte fertilization rate was not affected by TPLSM exposure, but the developmental capacity of the denuded oocytes was significantly reduced. This is, to our knowledge, the first article describing repeated intravital imaging during mammalian oocyte maturation using TPLSM.

scholarly journals PENGGUNAAN CAIRAN FOLIKEL DALAM MEDIA MATURASI IN VITRO OOSIT KAMBING BLIGON

2014 ◽  
Vol 8 (1) ◽  
Author(s):  
Diah Tri Widayati ◽  
Devita Hery Fatmawati ◽  
Nony Ariesta ◽  
Kustono K
Keyword(s):  
Tissue Culture ◽  
Culture Medium ◽  
Tissue Culture Medium ◽  
Phosphate Buffered Saline

Penelitian ini bertujuan mengetahui pengaruh medium maturasi terhadap kompetensi perkembangan in vitro oosit kambing bligon. Ovarium kambing diperoleh dari Rumah Potong Hewan, dibawa ke laboratorium dalam 0,9% NaCl pada suhu 31-34 C. Oosit diaspirasi dari folikel berdiameter 2-6 mm dengan menggunakan syringe 3 ml dan jarum 23 G. Oosit dengan minimal 2 lapisan sel-sel kumulus diproses untuk maturasi in vitro. Oosit dicuci 3 kali dengan dulbecco’s phosphate buffered saline (DPBS) dan tissue culture medium 199 (TCM) 199, kemudian didistribusikan ke dalam 50 μl medium maturasi yang berbeda, yaitu TCM 199 (tanpa suplementasi),

scholarly journals L-carnitine Supplementation Enhances Nuclear and Cytoplasmic Maturation Rates of Sheep Oocytes In Vitro

2021 ◽  
Vol 44 (2) ◽  
pp. 131-137
Author(s):  
Z. W. Bhakty ◽  
E. M. Kaiin ◽  
N. W. K. Karja ◽  
M. A. Setiadi
Keyword(s):  
Tissue Culture ◽  
Treatment Group ◽  
Culture Medium ◽  
Fertilization Rate ◽  
Control Group ◽  
Carnitine Supplementation ◽  
Progressive Motility ◽  
Maturation Medium ◽  
Cytoplasmic Maturation

The aim of the present study was to determine the effectiveness of l-carnitine (LC) supplementation on nuclear and cytoplasmic maturation rates of sheep oocytes. In experiment 1, oocytes were maturated for 24 hours in tissue culture medium 199 supplemented with LC at doses of 0.3 mg/mL, 0.6 mg/mL, and 0.9 mg/mL. In experiment 2, oocytes were maturated and fertilized in a media supplemented with LC at a dose of 0.3 mg/mL and incubated with 5x106 sperm/mL for 12 hours. The treatment group consisted of LC supplementation only in maturation medium (P1), only in fertilization medium (P2), and in both maturation and fertilization media (P3). In experiment 3, sperm motility patterns were assessed using CASA after being exposed to fertilization medium supplemented with LC at a dose of 0.3 mg/mL for 0 and 3 hours. Our results showed that supplementation of LC at a dose of 0.3 mg/mL significantly (p<0.05) increased the percentage of oocytes reaching metaphase II (86.7±4.1%) compared to those supplemented with LA at doses of 0, 0.6, and 0.9 mg/mL (73.6±1.2%, 81.4±1.3%, and 70.5±1.6%, respectively). The LC treatment in the fertilization medium only did not influence the number of two pronuclear formations (62.1±2.5%) compared to supplementation either in the maturation medium only (72.0±4.7%) or a combination of both in maturation and fertilization media (68.2±2.7%) (p<0.05). Further results after 3 hours of incubation compared to the control group showed the total motility (24.8±2.04% vs. 17.49±2.37%), progressive motility (14.17±2.03% vs. 6.49±1.64%), and curvilinear velocity (VCL) (119.70±3.73% vs. 71.15±10.59%) (p<0.05) were increased in the fertilization medium containing LC but it did not improve the fertilization rate. It is concluded that supplementation of LC at a dose of 0.3 mg/mL in the maturation medium only could better improve the nuclear and cytoplasmic maturation rates of sheep oocytes.

scholarly journals In Vitro Screening Ketahanan Galur Padi (Oryza Sativa) B7 Hasil Rakitan Politeknik Negeri Lampung Terhadap Keracunan Besi (Fe)

2019 ◽  
Vol 19 (3) ◽  
pp. 241
Author(s):  
Onny Chrisna Pandu Pradana ◽  
Siti Novridha Andini
Keyword(s):  
Tissue Culture ◽  
Oryza Sativa ◽  
Iron Toxicity ◽  
In Vitro Screening ◽  
Culture Bottle ◽  
Randomized Design ◽  
Significant Difference ◽  
Completely Randomized Design ◽  
And Control

This research aimed to invstigate the response of paddy culture (B7 strain) assembled by Lampung State Polytechnic to the iron toxicity tolerance. The research was done at Plant Tissue Culture Laboratory, Lampung State Polytechnic, from July to September 2019. Treatments were single arranged in a completely randomized design with three replications. The treatment tried was five levels of Fe concentrations (5,6 ppm 28 ppm, 56 ppm, 84 ppm, 112 ppm, and control). Each replication consisted of three culture bottle containing one explant. The homogenity of data was tested using Barlett test. If the assumption were fulfilled then analysis of variance is executed using STATISTIX 10, and then followed by the Honest Significant Difference (HSD) test in 5% alpha for mean separation and RPA analysis. The result of this research showed that the B7 strain has tolerance to iron toxicity until 56 ppm of Fe concentration, it can be concluded from the PAR value of its strain (>0,50). Meanwhile in  84 and 112 ppm of Fe concentration, the RPA value of B7 starin (<0,50), and it is indicate that its strain is sensitive. 

scholarly journals Feedback regulation of granulopoiesis: polymerization of lactoferrin abrogates its ability to inhibit CSA production

Blood ◽  
1982 ◽  
Vol 60 (1) ◽  
pp. 108-112 ◽  
Author(s):  
GC Jr Bagby ◽  
RM Bennett
Keyword(s):  
Tissue Culture ◽  
Culture Medium ◽  
Feedback Regulation ◽  
Tissue Culture Medium ◽  
Mononuclear Leukocytes ◽  
Gel Chromatography ◽  
Polymeric Form ◽  
Higher Doses

Neutrophil extracts were prepared from the peripheral blood of 40 normal volunteers and tested for their ability to inhibit CSA production by mononuclear leukocytes. Highly dilute neutrophil extracts inhibited CSA production/release, while extracts selectively depleted of lactoferrin by antibody affinity chromatography did not. In addition, higher concentrations of neutrophil extracts and higher doses of lactoferrin (10(-9)-10(-6) M) failed to inhibit CSA production/release. We found no evidence of CSA or CSA-enhancing factors in either our lactoferrin or our neutrophil extracts. However, using gel chromatography and rate zonal density sedimentation, we noted that lactoferrin undergoes concentration-dependent polymerization at 10(-9)-10(-10) M in tissue culture medium and that while monomeric lactoferrin effectively inhibits CSA production/release in vitro, the polymeric form does not. Thus, while we have confirmed that lactoferrin is the activity in neutrophil extracts that inhibits CSA production, we have also found that lactoferrin undergoes reversible polymerization at physiologic concentrations and that the polymerized molecule is inactive. The tendency of lactoferrin to polymerize in tissue culture medium and in vivo should be taken into account in any studies on its potential role as a physiologically relevant regulator of granulopoiesis.

scholarly journals Pertumbuhan Tunas Pisang Barangan (Musa Acuminata Colla.) akibat Pemberian Benzyl Amino Purin dan Arang Aktif secara In Vitro.

2019 ◽  
Vol 4 (1) ◽  
pp. 73-90
Author(s):  
Mulia Saputri ◽  
Marai Rahmawati ◽  
Elly Kesumawati
Keyword(s):  
Tissue Culture ◽  
Activated Charcoal ◽  
Shoot Growth ◽  
Growth Time ◽  
Shoot Height ◽  
High Productivity ◽  
Horticultural Crops ◽  
Randomized Design ◽  
And Control

Pertumbuhan Tunas Pisang Barangan Akibat Pemberian Benzyl AminoPurin dan Arang Aktif secara In Vitro Accretion of Barangan Banana Shoot Effect of  (BAP)  and Activated Charcoal Explant by In VitroAbstrak. Banana as a superior product of horticultural crops, has not achieved high productivity and has several obstacles in its multiplication. Tissue culture is one solution to overcome this problem. This study aims to determine the composition of PGRBenzil Amino Purine (BAP) and activated charcoal that are appropriate in the multiplication of barangan banana shoots. This research was conducted at the Tissue Culture Laboratory Faculty of Agriculture Syiah Kuala University, Darussalam Banda Aceh. The design used was a Completely Randomized Design (CRD) with two treatment factors. The first factor is BAP concentration consisting of 3 levels, namely 4 mg/L, 6 mg /L, and 8 mg/L. The second factor is activated charcoal concentration consisting of 3 levels, namely control, 1 g/L and 2 g/L. In this study, from 9 treatment combinations, only 4 treatment combinations were not contaminated. Of the 4 treatment combinations the combination of BAP concentration of 6 mg/L and control (without activated charcoal) showed the fastest shoot growth time of 29 days after multiplication, the most shoot growth was 6 shoots and the average shoot height was 15.9 mm

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