scholarly journals Binding of diethylstilboestrol to deoxyribonucleic acid by rat liver microsomal fractions in vitro and in mouse foetal cells in culture

1976 ◽  
Vol 158 (3) ◽  
pp. 643-646 ◽  
Author(s):  
G M Blackburn ◽  
M H Thompson ◽  
H W S King
Keyword(s):  
Tissue Culture ◽  
Culture System ◽  
Deoxyribonucleic Acid ◽  
Microsomal Preparation ◽  
Enzymic Digestion ◽  
Cells In Culture ◽  
Primary Mouse ◽  
Significant Yield ◽  
Liver Microsomal Preparation

Diethylstillboestrol, a synthetic and carcinogenic hormone, binds to DNA as a result of incubation with a liver microsomal preparation in vitro and on incubation with primary mouse foetal cells in culture. Enzymic digestion of DNA samples thus prepared gives several covalent deoxyribonucleoside-diethylstilboestrol products from the microsomal system. One of these is produced in small but significant yield in the tissue-culture system.

Cultured epithelial cells derived from human foetal pancreas as a model for the study of cystic fibrosis: further analyses on the origins and nature of the cell types

1988 ◽  
Vol 90 (1) ◽  
pp. 73-77
Author(s):  
A. Harris ◽  
L. Coleman
Keyword(s):  
Cystic Fibrosis ◽  
Tissue Culture ◽  
Epithelial Cells ◽  
Culture System ◽  
Cultured Cells ◽  
Cell Types ◽  
Cultured Epithelial Cells ◽  
Different Cell Types ◽  
Foetal Pancreas

The establishment of a tissue-culture system for epithelial cells derived from human foetal pancreas has recently been reported. Further analyses have now been made on these cells in vitro, together with parallel investigation of the distribution of different cell types within the intact foetal pancreas. Results support the view that the cultured cells are ductal in origin and nature. Pancreatic epithelial cell cultures have also been established from foetuses with cystic fibrosis.

scholarly journals 213.Culture of mouse male germ cells for genetic manipulations

2004 ◽  
Vol 16 (9) ◽  
pp. 213
Author(s):  
C. M. Itman ◽  
B. Barakat ◽  
K. Loveland
Keyword(s):  
Gene Transfer ◽  
Germ Cell ◽  
Germ Cells ◽  
Cell Biology ◽  
Culture System ◽  
Striking Difference ◽  
Embryonal Carcinoma Cell ◽  
Cells In Culture ◽  
New Genes

The ability to maintain germ cells in culture offers the opportunity to manipulate their differentiation and to deliver new genes through the germline. However the study of male germ cell biology is hindered by the lack of an in vitro culture system that supports germ cell maintenance and differentiation, and the lack of efficient gene transfer technologies. To address this, we developed a short term (1-6 day) in vitro germ cell-Sertoli cell co-culture system using testes of 6 day old mice, in which the only germ cell type present is the spermatogonium. To establish culture conditions favourable for maintaining germ cells in culture, we compared different substrates (plastic, laminin, poly-L-lysine, Matrigel) and cell culture additives (insulin/transferrin, retinoic acid, lactic acid, pyruvic acid). Using immunocytochemistry, we observed that germ cell numbers varied depending on the substrate, and was greatly improved on laminin or Matrigel. The culture additives tested had no effect. We noted a striking difference in germ cell growth depending on the mouse strain used, with C57BlXCBA F1 cross > Swiss > BalbC >> C57Bl/6J. To investigate gene transfer into germ cells, constructs were generated using promoters identified as exhibiting germ cell-specific expression in transgenic animals (EF1-α, Oct4, Stra8) with EGFP and lacz reporter genes. From this baseline, we have started gene transfer experiments. Initial transfection experiments were performed in mouse testis cell lines (GC1, GC2, TM3, TM4) and the P19 mouse embryonal carcinoma cell line. Several commercial transfection agents and a non-commercial product (1) were used, as was electroporation, however these did not yield reporter expression in germ cells. As these techniques require proliferating cells, we are now undertaking a study with the addition of growth factors in culture (BMP4, activin, GDNF, FSH) to enhance germ cell proliferation and therefore improve transfection efficiencies. We will also employ lentiviral transduction, as this is reportedly 100% efficient and independent of proliferation. (1) Celebi et al. (2002) Mol. Reprod. Dev. 62,477–482.

Enzymic Cleavage Of In Vivo Formed Maillard-Type Compounds Involved In Haemostasis

1981 ◽  
Author(s):  
L Mester ◽  
L Szabados ◽  
M Mester
Keyword(s):  
Enzyme System ◽  
Basal Membrane ◽  
Enzyme Concentration ◽  
Cytochrome P450 Enzyme ◽  
Microsomal Preparation ◽  
Chemical Hydrolysis ◽  
P450 Enzyme ◽  
Liver Microsomal Preparation

Desoxyfructose derivatives of serotonin (Mester et al.,1975), of haemoglobin (Flückiger and Winterhalter, 1976), of poly-L-lysine (Mester et al., 1975) and of lysine rich histones (Kertesz-Crisba, 1977) are easily formed in vivo by a simple Maillard-type chemical reaction. Some of these compounds interfere with platelet functions (Mester et al.,1976) or contribute to the thickening of the basal membrane of blood vessels (Cerami et al., 1979).While the chemical synthesis of Maillard-type compounds proceeds readily even in vivo, the chemical cleavage of them needs sever conditions which certainly do not exist in vivo (Gottschalk, 1952). However, a slow liberation of serotonin from desoxyfructo-serotonin is observed in vivo, suggesting the existence of an enzyme system for the cleavage of Maillard-type sugar-amine derivatives. In vitro, using a sheep liver microsomal preparation rich in Cytochrome P450 enzyme, the liberation of serotonin is in linear correlation with the enzyme concentration. The cleavage of desoxyfructo-serotonin is activated by NADPH having its optimum at pH=7.4, excluding definitely the occurence of a chemical hydrolysis.Factors interfering with the enzyme system involved in the cleavage of Maillard-type compounds, may also interfere with haemostasis.

Improved in vitro micropropagation method with adventitious corms and roots for endangered saffron

2012 ◽  
Vol 7 (1) ◽  
pp. 138-145 ◽  
Author(s):  
Evrim Zeybek ◽  
Sertaç Önde ◽  
Zeki Kaya
Keyword(s):  
Tissue Culture ◽  
Ascorbic Acid ◽  
Culture System ◽  
Crocus Sativus ◽  
Ex Situ ◽  
Root Initiation ◽  
Indirect Organogenesis ◽  
Adventitious Shoot Formation ◽  
Tissue Culture System

AbstractThe objective of this study was to investigate development of an efficient in vitro tissue culture system for saffron (Crocus sativus L.) complete with roots and corms. In indirect organogenesis, Murashige and Skoog (MS) media with 3% (w/v) sucrose, 100 mg L−1 ascorbic acid, and the combination of 0.25 mg L−1 2,4-dichlorophenoxyacetic acid (2,4-D) and 1 mg L−1 6-benzylaminopurine (BAP) were best for callus initiation and growth while 1.5 mg L−1 BAP was excellent for high rate of adventitious shoot formation. 1 mg L−1 indole-3-butyric acid (IBA) was more preferable for adventitious corm and root initiation as well as growth. Overall, 64% rooting and 33% corm production rates were achieved in indirect organogenesis. In direct organogenesis, MS medium supplemented with 3 % sucrose, 100 mg L−1 ascorbic acid and 1 mg L−1 BAP was optimum for shoot growth. While 1 mg L−1 IBA was best for adventitious corm formation, 2 mg L−1 IBA promoted adventitious root initiation and growth. Overall, 36% and 57% of explants had corm and contractile root, respectively. The high rates suggest that efficient tissue culture system could be achieved for mass propagation and ex situ conservation of threatened saffron genetic resources.

scholarly journals The Porcine Aortic Tissue Culture System in vitro for Stem Cell Research

2011 ◽  
Vol 4 (2) ◽  
pp. 116-122
Author(s):  
Dong-Eun Kim ◽  
Keun-Hee Oh ◽  
Ji-Hye Yang ◽  
Sun-Keun Kwon ◽  
Tae-Jun Cho ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Stem Cell ◽  
Stem Cell Research ◽  
Culture System ◽  
Aortic Tissue ◽  
Tissue Culture System

Establishment of a tissue culture system for epithelial cells derived from human pancreas: a model for the study of cystic fibrosis

1987 ◽  
Vol 87 (5) ◽  
pp. 695-703
Author(s):  
A. Harris ◽  
L. Coleman
Keyword(s):  
Cystic Fibrosis ◽  
Tissue Culture ◽  
Epithelial Cells ◽  
Culture System ◽  
Cultured Cells ◽  
Cell Types ◽  
Human Pancreas ◽  
Tissue Culture System ◽  
Molecular Biological Analysis

A tissue culture system for epithelial cells derived from human foetal pancreas has been established. The cultured cells show many ultrastructural features of interlobular duct cells. Immunocytochemical and histochemical evidence is presented in support of the view that these cells are ductal in origin. They are likely to be one of the few cell types that express the basic defect of cystic fibrosis in vitro. The cells may be passaged and sufficient material obtained to permit biochemical and molecular biological analysis.

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