Factors Affecting Rainbow Trout Ovary Cells Cultivated In vitro

1969 ◽  
Vol 26 (2) ◽  
pp. 461-463 ◽  
Author(s):  
M. F. Li ◽  
Carol Jordan

The rainbow trout ovary cells proliferated well at 18 and 23 C, and at culture medium pH value of 7.1 and 7.9. Outside these limits cell proliferation was hindered and the cellular morphology was affected significantly. The ovary cells have been shown to be susceptible to amphibian virus (LT-1).

2021 ◽  
Vol 843 (1) ◽  
pp. 012041
Author(s):  
A A Verbitskaya ◽  
A S Ivanova ◽  
N G Konkova ◽  
A K Gaponenko

Abstract The aim of this research was to study the morphogenetic ability of Taraxacum kok-saghyz root tissues and to optimize the culture medium for the subsequent genetic transformation of plants. The effects of exogenous hormone exposure on survival, in vitro shoot induction, and root formation were studied by using root tissues. For the cultivation the samples of kok-saghyz, the Murashige-Skuga nutrient medium was used as the basis, supplemented with sucrose 20 g/l, vitamins B5 1 mg/l, and also containing agar 5.5 g/l. The pH value is 5.7. For the plant regeneration induction, growth regulators, auxins and cytokinins were added in culture media. In this study, the roots of D 1 mm and two media variants were used for comparison: variant 1. MS + 6-BAP 1 mg/l; variant 2. MS + 6-BAP 1 mg/l + IAA 0.2 mg/l. An effective protocol for the regeneration of kok-saghyz explants was developed. There was a high percentage of regeneration of 87.6% on the medium containing a combination of cytokinin and auxin, as well as a high percentage of direct shoot formation, which was 65.1%, the degree of rooting was 100%, the resulting cultured plant tissues grew well and had a high survival rate after transplantation.


1988 ◽  
Vol 255 (3) ◽  
pp. H554-H562 ◽  
Author(s):  
C. J. Meininger ◽  
M. E. Schelling ◽  
H. J. Granger

The proliferation of bovine aortic or coronary venular endothelial cells (EC) in vitro was stimulated by the addition of adenosine (0.5 or 5.0 microM) to the culture medium. Cell counts of adenosine-treated aortic EC were 23–76% and coronary venular EC 19–52% greater than nontreated controls. Because adenosine is known to be released by hypoxic tissues, cell proliferation was quantitated when aortic EC were grown at 2% O2. Cell counts were 41–102% greater under hypoxic conditions than when cells were grown at standard tissue culture conditions (approximately 20% O2). When culture medium conditioned by coronary EC grown at 2% O2 was added to EC growing at standard conditions, cell counts were 24–69% greater than controls with medium conditioned by coronary EC grown at 20% O2. This suggests that hypoxia causes endothelial cells to release a factor(s) into the medium that can stimulate cell proliferation. The addition of the adenosine receptor blocker 8-phenyltheophylline (10(-5) M) prevented the stimulation of proliferation caused by hypoxia-conditioned medium, 2% O2 or 5.0 microM adenosine, suggesting that adenosine mediates its effect via an external membrane receptor. Adenosine also stimulated EC chemotaxis. Taken together, these results suggest that adenosine, released as a result of tissue hypoxia, may act as an angiogenic stimulus for the growth of new blood vessels.


2004 ◽  
Vol 286 (5) ◽  
pp. R935-R941 ◽  
Author(s):  
Juan Castillo ◽  
Marta Codina ◽  
María Laura Martínez ◽  
Isabel Navarro ◽  
Joaquim Gutiérrez

The relative function of IGF-I and insulin on fish muscle metabolism and growth has been investigated by the isolation and culture at different stages (myoblasts at day 1, myocytes at day 4, and myotubes at day 10) of rainbow trout muscle cells. This in vitro model avoids interactions with endogenous peptides, which could interfere with the muscle response. In these cells, the effects of IGF-I and insulin on cell proliferation, 2-deoxyglucose (2-DG), and l-alanine uptake at different development stages, and the use of inhibitors were studied and quantified. Insulin (10-1,000 nM) and IGF-I (10-100 nM) stimulated 2-DG uptake in trout myocytes at day 4 in a similar manner (maximum of 124% for insulin and of 142% for IGF-I), and this stimulation increased when cells differentiated to myotubes (maximum for IGF-I of 193%). When incubating the cells with PD-98059 and especially cytochalasin B, a reduction in 2-DG uptake was observed, suggesting that glucose transport takes place through specific facilitative transporters. IGF-I (1-100 nM) stimulated the l-alanine uptake in myocytes at day 4 (maximum of 239%), reaching higher values of stimulation than insulin (100-1,000 nM) (maximum of 160%). This stimulation decreased when cells developed to myotubes at day 10 (118% for IGF-I and 114% for insulin). IGF-I (0.125-25 nM) had a significant effect on myoblast proliferation, measured by thymidine incorporation (maximum of 170%), and required the presence of 2-5% fetal serum (FBS) to promote thymidine uptake. On the other hand, insulin was totally ineffective in stimulating thymidine uptake. We conclude that IGF-I is more effective than insulin in stimulating glucose and alanine uptake in rainbow trout myosatellite cells and that the degree of stimulation changes when cells differentiate to myotubes. IGF-I stimulates cell proliferation in this model of muscle in vitro and insulin does not. These results indicate the important role of IGF-I on growth and metabolism of fish muscle.


1995 ◽  
Vol 62 (4) ◽  
pp. 577-586 ◽  
Author(s):  
Boris Zavizion ◽  
A. John Bramley ◽  
Ioannis Politis

SUMMARYThe effects of culture supernatants conditioned by the growth ofStaphylococcus aureusM60 onin vitrogrowth and functional properties of bovine mammary myoepithelial cells were examined. Myoepithelial cell proliferation was reduced byStaph, aurexisM60 culture supernatants. Exposure of myoepithelial cells to culture supernatants of isogeneic mutants ofStaph, aureusM60 that produced either α or β toxins reduced proliferation, but to a lesser extent than supernatants from the wild type strain. Thus, α and β toxins may play some role in affecting myoepithelial cell proliferation. Of the cells tested, 42% contracted following addition of oxytocin (10–7M) in the culture medium. Treatment of myoepithelial cells for 15 min withStaph, aureusM60 supernatants, prior to addition of oxytocin in the culture medium, increased the number of cells that contracted to 92%. Exposure of cells for 3 h to the same supernatant, prior to addition of oxytocin in the culture medium, abolished oxytocin responsiveness, had no effect on immunolocalization of actin and vimentin, but affected the localization of α-actinin within myoepithelial cells. Treatment of myoepithelial cells for 3 h with a combination of purified staphylococcal proteinases XVI and XVII-B abolished oxytocin responsiveness and mimicked the effect of theStaph, aureusculture supernatant. We conclude thatStaph, aureusM60 culture supernatant affected proliferation and functional properties of myoepithelial cells.


1997 ◽  
Vol 75 (9) ◽  
pp. 1474-1478 ◽  
Author(s):  
Mary J. Packard

Factors affecting release of calcium by the chick chorioallantoic membrane in vitro were assessed by culturing explants of shell with and without the chorioallantoic epithelium. Explants were removed from eggs on day 16 of incubation and cultured using techniques similar to those used to examine resorption of bone in vitro. The region of an egg from which an explant is removed, the strain of donor eggs used, and addition of bovine serum albumin to the culture medium did not affect release of calcium. In contrast, exposure of explants to a variety of different culture media (DMEM, Medium 199, BGJb, and RPMI) introduced considerable variation in calcium release. Calcium release in vitro was also examined by removing the blunt end of a fertile egg as well as the embryo and yolk. The remaining preparation was filled with culture medium, covered, and cultured for two 24-h periods, with a change of medium after the first 24 h. Preparations from which the chorioallantoic membrane had been removed released very little calcium during culture, whereas those with the epithelium in place released considerable quantities. However, release of calcium by preparations with the membrane declined during the second interval. When surface area is taken into account, such preparations mobilize less calcium than explants cultured under similar conditions.


2002 ◽  
Vol 60 (1-2) ◽  
pp. 43-59 ◽  
Author(s):  
Michael Kilemade ◽  
Maria Lyons-Alcantara ◽  
Tina Rose ◽  
Richard Fitzgerald ◽  
Carmel Mothersill

2021 ◽  
Vol 9 (Suppl 3) ◽  
pp. A639-A639
Author(s):  
Feng Jiang ◽  
Hong Wang ◽  
Tsz-Lun Yeung ◽  
Guozhong Qin ◽  
Bo Marelli ◽  
...  

BackgroundBintrafusp alfa is a first-in-class bifunctional fusion protein composed of the extracellular domain of the TGF-βRII receptor fused to a human IgG1 antibody blocking PD-L1. The TGF-βRII moiety of bintrafusp alfa functions as a ”trap” to sequester active TGF-β but does not block TGF-β release from its latent form. Multiple mechanisms lead to the release of active TGF-β. Integrins control local activation of latent TGF-β stored in the extracellular matrix and cell-surface reservoirs in the tumor microenvironment (TME). Alpha v integrin mRNA expression is correlated with multiple TGF-β gene signatures. It has been shown that αvβ8 integrin mediates TGF-β activation without releasing it from the latent TGF-β complex, suggesting that the TGF-βRII moiety of bintrafusp alfa may be unable to sequester TGF-β activated by αvβ8 integrin. Therefore, we hypothesize that combining abituzumab, a pan–αv integrin antibody, with bintrafusp alfa may lead to enhanced suppression of TGF-β signaling.MethodsThe expression of αv and β6 integrin mRNA was determined by RNA sequencing of triple-negative breast cancer (TNBC) tumor samples from a phase 1 clinical trial of bintrafusp alfa and correlated with patient response to bintrafusp alfa. The combination of bintrafusp alfa and abituzumab was investigated in vitro and in vivo in a TGF-β–dependent human tumor model, Detroit 562. In this study, CellTiter-Glo 2.0 Assay measured cell proliferation in vitro and enzyme-linked immunosorbent assay measured the level of latency-associated protein (LAP). A TGF-β reporter cell line MDA-MB-231 measured the level of active TGF-β. Antitumor activity in vivo was evaluated via tumor growth of Detroit 562 xenograft model in SCID mice.ResultsIn TNBC, increased expression of αv and β6 integrin mRNA was associated with poor response to bintrafusp alfa, suggesting that TGF-β activated by αv integrin may not be blocked by bintrafusp alfa. In Detroit 562 cells, abituzumab increased LAP levels in the cell culture medium, confirming modulation of the TGF-β pathway. As a result, the amount of active TGF-β released into culture medium was reduced by abituzumab. In vitro, both abituzumab and bintrafusp alfa suppressed Detroit 562 cell proliferation, and the combination suppressed cell proliferation further. In vivo, the combination led to increased tumor growth inhibition of Detroit 562 xenograft tumors relative to either monotherapy, further supporting the potential of this combination.ConclusionsCollectively, these preclinical findings support clinical development of bintrafusp alfa and abituzumab combination therapy to maximally suppress TGF-β signaling in the TME.AcknowledgementsWe thank George Locke for his analysis of the RNAseq data.Ethics ApprovalThis study was approved by the Institutional Animal Care and Use Committee at EMD Serono, Inc.; approval number [17–008].


2021 ◽  
Vol 9 (33) ◽  
pp. 147-155
Author(s):  
Solange Monteiro de Toledo Piza Gomes Carneiro ◽  
Euclides Davidson Bueno Romano ◽  
Erika Pignoni ◽  
Marcus Zulian Teixeira ◽  
Maria Elizabeth da Costa Vasconcelos ◽  
...  

Background: homeopathy is a means permitted in organic agriculture to control disease and plagues; biotherapics are a practical means for farmers to intervene on the health of plants in agro-ecological systems of production. Tomato-plants can be affected by several diseases, one of the most significant ones in Brazil is early blight, caused by fungus Alternaria solani, due to the damage it causes and its wide distribution in the country. Aims: to establish whether a biotherapic of A. solani may interfere on the in vitro development of the fungus and whether it affects the severity of early blight on tomato-plants in greenhouse. Methods: the effect of the biotherapic on the fungus was evaluated through the percentage of germinated spores under microscope and the growth of colonies in a culture medium. Treatments used were: biotherapic 26cH, 27cH, 28cH, 29cH and 30cH; sterilized distilled water; and diluted and agitated hydroalcoholic solution. The effect of the biotherapic on the development of disease was evaluated in 4 experiments in greenhouse. Plants were kept in vases and subjected to artificial inoculation of the fungus after the application of treatments. Evaluation of disease was carried out through diagrammatic scale. Results: no treatment affected the germination of spores or the development of fungus colonies in the culture medium. In the first test, treatment 26cH differed from water in Tukey’s test at 5% but did not differed from diluted and agitated hydroalcoholic solution. In the second test, treatments 27cH and 28cH showed significant difference from both water and hydroalcoholic solution with an average control of disease of 57% and 62% respectively. The other 2 tests did nor exhibit any significant effect. Conclusions: there was no direct effect of the biotherapic on the fungus, but there was an effect on the severity of the disease. Factors affecting the efficiency of the biotherapic must be better understood before it can be recommended to farmers for the management of early blight in tomato-plants.


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