Use of a tissue culture medium for in vitro studies on the ion transport capacity of amphibian epithelia

1975 ◽  
Vol 31 (12) ◽  
pp. 1418-1420 ◽  
Author(s):  
M. G. Emílio ◽  
M. Balls ◽  
H. Menano
Keyword(s):  
Tissue Culture ◽  
Ion Transport ◽  
Culture Medium ◽  
In Vitro Studies ◽  
Tissue Culture Medium ◽  
Transport Capacity ◽  
Amphibian Epithelia

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 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 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.

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 In vitro Fertilization in Iraqi Local Goats

2009 ◽  
Vol 3 (1) ◽  
pp. 5-9
Author(s):  
Hazim I. AL-Ahmed
Keyword(s):  
Tissue Culture ◽  
In Vitro Fertilization ◽  
Culture Medium ◽  
Polar Body ◽  
Culture Technique ◽  
Tissue Culture Medium ◽  
Vitro Fertilization ◽  
Cumulus Oophorus ◽  
First Polar Body

412 primary ova were used in the study of in vitro fertilization, these ova were collected from ovaries samples of does at different stages of oestrous cycle collected from abattoirs.These follicles were classified according to their size into large follicles (> 2-6 mm) and small follicles (1-2 mm). Ova aspirated from these follicles were evaluated depending on the presence or absence of cumulus oophorus cells and on the presence of the first polar body. The aspirated ova from large and small follicles were maturated in tissue culture medium 199 to study their ability of maturation.. The microdrops technique from tissue culture (Medium 199) and granulosa cell co-culture technique were used for the maturation of ova, also the in vitro fertilization was inducted in these ova with the sperm which were capacitated in Bracket Medium. The results showed that the highest rate for ova aspiration and the highest rate of ova surrounded by cumulas oophorus were from the large, follicles. The size of follicle has a significant influence on the degree of ova growth and maturation. The results showed the absence of significant differences in the efficacy of two techniques used in the ova maturation and their ability of fertilization.

scholarly journals Criopreservação de embriões de suínos adicionando trealose aos crioprotetores etilenoglicol ou glicerol

1999 ◽  
Vol 29 (1) ◽  
pp. 111-115
Author(s):  
Edmir da Silva Nicola ◽  
João Carlos Deschamps ◽  
Milton Macedo Júnior ◽  
José Bozato Sobrinho
Keyword(s):  
Tissue Culture ◽  
Culture Medium ◽  
Tissue Culture Medium

Os objetivos deste trabalho foram: comparar (a) o efeito da adição de trealose ao crioprotetor etilenoglicol e, (b) o efeito dos crioprotetores etilenoglicol ou glicerol associados à trealose sobre a viabilidade de embriões congelados de suínos . A trealose foi adicionada à solução de congelamento de etilenoglicol 1.5 M nas concentrações de 0.0 M (tratamento 1) e 0.25 M (tratamento 2), e na concentração de 0.25 M (tratamento 3) à solução de congelamento de glicerol 1.5 M. Os embriões foram congelados no estágio de blastocisto expandido. O método de congelamento utilizado foi o rápido, sendo que desde a temperatura ambiente (± 25°C) até a temperatura do "seeding" (-7°C), a velocidade de resfriamento foi de 1°C/minuto. Após o "seeding", foi estabelecido um período de equilíbrio de 10 minutos à -7°C, sendo, a partir de então, adotada a velocidade de resfriamento de 0.3°C/minuto até a temperatura de -35°C. A seguir, as palhetas foram colocadas no nitrogênio líquido (-196°C). O descongelamento foi realizado mediante exposição das palhetas ao ar durante 30 segundos e, posteriormente, em banho-maria à 35°C, durante 30 segundos. A remoção dos crioprotetores foi realizada pelo método em etapas. O critério de viabilidade adotado consistiu na observação da reexpansão da cavidade blastocélica dos embriões, após período de cultivo in vitro de 18-24h. O cultivo foi feito em meio TCM 199 (tissue culture medium), acrescido de 20% de soro fetal bovino, em microgotas de 60 mil, sob óleo mineral em estufa com 6% CO2, 95% de umidade e 37°C. A viabilidade embrionária logo após o descongelamento (0h) foi de 17.2%, 37.5% e 42.8%, para os tratamentos 1, 2 e 3, respectivamente. Após o período de cultivo de 18-24h, a viabilidade embrionária foi de 6.9%, 28.1% e 28.5%, para os tratamentos 1, 2 e 3, respectivamente. Os resultados indicaram que não houve diferença (p>0.05) significativa entre os tratamentos ao descongelamento. Após o cultivo in vitro, verificou-se que a porcentagem de embriões viáveis foi superior (p<0.05) pela adição de trealose na concentração de 0.25 M à solução de congelamento, contendo etilenoglicol. No entanto, não foi detectada diferença significativa entre os crioprotetores etilenoglicol ou glicerol, associados à trealose.

scholarly journals Comparison of Quinn's Advantage fertilization medium and tissue culture medium 199 for in vitro maturation of oocytes

2014 ◽  
Vol 53 (1) ◽  
pp. 17-20 ◽  
Author(s):  
Yu-Hung Lin ◽  
Jiann-Loung Hwang ◽  
Lee-Wen Huang ◽  
Kok-Min Seow ◽  
Bih-Chwen Hsieh ◽  
...  
Keyword(s):  
Tissue Culture ◽  
In Vitro Maturation ◽  
Culture Medium ◽  
Tissue Culture Medium
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